Existing Annotations Review

GO Term	Evidence	Action	Reason
GO:0016887 ATP hydrolysis activity	IBA GO_REF:0000033	ACCEPT	Summary: Phylogenetic annotation of GET3's ATP hydrolysis activity, the catalytic core of its targeting cycle. Conserved across the ArsA/Get3 family. Reason: Core molecular function; GET3 is an ATPase whose ATP hydrolysis drives TA-protein release/insertion, supported by IDA and EC 3.6.4.-. Supporting Evidence: file:human/GET3/GET3-uniprot.txt ATP hydrolysis is required for insertion.
GO:0071816 tail-anchored membrane protein insertion into ER membrane	IBA GO_REF:0000033	ACCEPT	Summary: Phylogenetic annotation of GET3's defining biological process. This is the standard GO term used for the GET-pathway role; GET3 is the cytosolic targeting factor whose action commits TA proteins to ER insertion. Reason: Core biological process; conserved and supported by experimental evidence (IMP/IDA). Supporting Evidence: file:human/GET3/GET3-uniprot.txt ATPase required for the post-translational delivery of tail-
GO:0005524 ATP binding	IEA GO_REF:0000120	KEEP AS NON CORE	Summary: ATP binding drives the GET3 homodimer to the closed state that captures the TA substrate. A structural/mechanistic attribute subsidiary to the catalytic ATP hydrolysis activity. Reason: Accurate (GET3 binds ATP) but subsidiary to the more informative ATP hydrolysis activity that represents the core MF. Supporting Evidence: file:human/GET3/GET3-uniprot.txt ATP binding drives the
GO:0005730 nucleolus	IEA GO_REF:0000044	KEEP AS NON CORE	Summary: Electronic transfer of a legacy nucleolar localization reported in early arsenite-ATPase studies. Not tied to the core cytosolic TA-targeting function. Reason: Legacy/secondary localization derived from early arsA-homolog studies; not part of the core GET-pathway function. Supporting Evidence: file:human/GET3/GET3-uniprot.txt Nucleus, nucleolus
GO:0005737 cytoplasm	IEA GO_REF:0000044	ACCEPT	Summary: Electronic transfer of the cytoplasmic localization, the primary site where GET3 captures TA substrates. Consistent with experimental evidence. Reason: Correct primary compartment; GET3 is a cytosolic targeting factor. Supporting Evidence: file:human/GET3/GET3-uniprot.txt SUBCELLULAR LOCATION: Cytoplasm
GO:0005783 endoplasmic reticulum	IEA GO_REF:0000120	ACCEPT	Summary: Electronic annotation of ER localization, where GET3 transiently docks on the WRB/CAML receptor to deliver TA substrates. Consistent with experimental evidence. Reason: Correct; GET3 visits the ER membrane to hand off its TA cargo. Supporting Evidence: file:human/GET3/GET3-uniprot.txt Endoplasmic
GO:0016887 ATP hydrolysis activity	IEA GO_REF:0000120	ACCEPT	Summary: Electronic assignment of the core ATP hydrolysis activity, consistent with experimental IDA and the catalytic activity record. Reason: Correct core molecular function; redundant with IDA/IBA. Supporting Evidence: file:human/GET3/GET3-uniprot.txt ATP hydrolysis is required for insertion.
GO:0045048 protein insertion into ER membrane	IEA GO_REF:0000104	KEEP AS NON CORE	Summary: Electronic (Swiss-Prot keyword/feature transfer) annotation of the parent ER protein-insertion process. Correct but less specific than the TA-insertion term. Reason: Correct but generic parent of GO:0071816, which better captures GET3's role. Supporting Evidence: file:human/GET3/GET3-uniprot.txt ATPase required for the post-translational delivery of tail-
GO:0005515 protein binding	IPI PMID:21516116 Next-generation sequencing to generate interactome datasets.	KEEP AS NON CORE	Summary: High-throughput interactome screen capturing GET3 protein interactions. The bare protein binding term is uninformative. Reason: Bare protein binding from a high-throughput screen; uninformative for the core MF. Supporting Evidence: PMID:21516116 Next-generation sequencing to generate interactome datasets
GO:0005515 protein binding	IPI PMID:21911467 Insight into bacterial virulence mechanisms against host imm...	KEEP AS NON CORE	Summary: Yersinia pestis-human protein-protein interaction screen; the captured partner is a bacterial xenobiotic protein (yscD), an incidental cross-species interaction unrelated to GET3's function. Reason: Incidental xenobiotic (bacterial) interaction from a host-pathogen screen; bare protein binding, not relevant to core function. Supporting Evidence: file:human/GET3/GET3-uniprot.txt O43681; Q56975: yscD
GO:0005515 protein binding	IPI PMID:25416956 A proteome-scale map of the human interactome network.	KEEP AS NON CORE	Summary: Proteome-scale human interactome map; source of IntAct partners including ER/secretory and TA-like proteins. Bare protein binding is uninformative. Reason: High-throughput interactome; bare protein binding term is uninformative. Supporting Evidence: PMID:25416956 A proteome-scale map of the human interactome network
GO:0005515 protein binding	IPI PMID:28514442 Architecture of the human interactome defines protein commun...	KEEP AS NON CORE	Summary: Human interactome architecture study; captures GET3 interactions including the functionally relevant CAMLG partner, but uses the uninformative bare protein binding term. Reason: Records real interactions (including CAMLG) but bare protein binding is uninformative; functional partners are captured via GET complex membership. Supporting Evidence: file:human/GET3/GET3-uniprot.txt O43681; P49069: CAMLG
GO:0005515 protein binding	IPI PMID:32296183 A reference map of the human binary protein interactome.	KEEP AS NON CORE	Summary: HuRI binary interactome (Y2H); source of many IntAct partners of GET3, including ER/secretory and TA-like proteins consistent with TA-client capture. Bare protein binding is uninformative. Reason: High-throughput Y2H interactome; bare protein binding is uninformative. Supporting Evidence: PMID:32296183 A reference map of the human binary protein interactome
GO:0005515 protein binding	IPI PMID:33961781 Dual proteome-scale networks reveal cell-specific remodeling...	KEEP AS NON CORE	Summary: Proteome-scale cell-specific interactome network; captures GET3 interactions including CAMLG. Bare protein binding is uninformative. Reason: High-throughput interactome; bare protein binding is uninformative. Supporting Evidence: file:human/GET3/GET3-uniprot.txt O43681; P49069: CAMLG
GO:0005515 protein binding	IPI PMID:40205054 Multimodal cell maps as a foundation for structural and func...	KEEP AS NON CORE	Summary: Multimodal cell map study; captures GET3 protein interactions including CAMLG. Bare protein binding is uninformative. Reason: High-throughput interactome; bare protein binding is uninformative. Supporting Evidence: file:human/GET3/GET3-uniprot.txt O43681; P49069: CAMLG
GO:0043529 GET complex	IEA GO_REF:0000120	ACCEPT	Summary: Electronic assignment of GET complex membership. GET3/TRC40 is a defining subunit of the GET complex (GET1/WRB + CAMLG/GET2 + GET3/TRC40). Reason: Core cellular component; consistent with experimental IDA/IPI evidence. Supporting Evidence: file:human/GET3/GET3-uniprot.txt Component of the Golgi to ER
GO:0071816 tail-anchored membrane protein insertion into ER membrane	IEA GO_REF:0000120	ACCEPT	Summary: Electronic assignment of the core TA-insertion process, consistent with experimental evidence. Reason: Correct core process; redundant with IMP/IDA/IBA. Supporting Evidence: file:human/GET3/GET3-uniprot.txt ATPase required for the post-translational delivery of tail-
GO:0005654 nucleoplasm	IDA GO_REF:0000052	KEEP AS NON CORE	Summary: HPA immunofluorescence nucleoplasm localization. A secondary localization not connected to the core cytosolic TA-targeting function. Reason: HPA-derived secondary localization; not part of the core GET-pathway function. Supporting Evidence: file:human/GET3/GET3-uniprot.txt Nucleus, nucleolus
GO:0005730 nucleolus	IDA GO_REF:0000052	KEEP AS NON CORE	Summary: HPA immunofluorescence nucleolar localization, consistent with the legacy nucleolar distribution reported for the arsenite-ATPase. Secondary to the core function. Reason: Secondary/legacy localization; not part of the core cytosolic targeting function. Supporting Evidence: file:human/GET3/GET3-uniprot.txt Nucleus, nucleolus
GO:0005737 cytoplasm	EXP PMID:17382883 Identification of a targeting factor for posttranslational m...	ACCEPT	Summary: Direct experimental cytoplasmic localization from the study that identified TRC40/Asna-1 as the cytosolic TA-targeting ATPase. Reason: Core compartment; GET3 acts as a cytosolic targeting factor. Supporting Evidence: PMID:17382883 cytosolic TMD recognition complex (TRC) that targets TA proteins for insertion into the ER membrane
GO:0005737 cytoplasm	EXP PMID:21444755 WRB is the receptor for TRC40/Asna1-mediated insertion of ta...	ACCEPT	Summary: Direct experimental cytoplasmic localization, consistent with GET3/TRC40 being a conserved cytosolic ATPase. Reason: Core compartment; corroborated by multiple experimental sources. Supporting Evidence: PMID:21444755 TRC40/Asna1 (Get3 in yeast)
GO:0005783 endoplasmic reticulum	EXP PMID:17382883 Identification of a targeting factor for posttranslational m...	ACCEPT	Summary: Direct experimental ER localization; GET3 docks on the ER membrane to deliver its TA cargo to the WRB/CAML receptor. Reason: Correct; GET3 transiently associates with the ER during TA delivery. Supporting Evidence: PMID:17382883 targets TA proteins for insertion into the ER membrane
GO:0005783 endoplasmic reticulum	EXP PMID:21444755 WRB is the receptor for TRC40/Asna1-mediated insertion of ta...	ACCEPT	Summary: Direct experimental ER localization, consistent with GET3 delivering TA proteins to the ER membrane receptor. Reason: Correct; GET3 visits the ER to hand off TA cargo. Supporting Evidence: PMID:21444755 delivers them to the ER membrane for insertion
GO:0005783 endoplasmic reticulum	EXP PMID:31461301 Biallelic Variants in ASNA1, Encoding a Cytosolic Targeting ...	ACCEPT	Summary: Experimental ER localization from the ASNA1 cardiomyopathy study, in which ASNA1 mediates TA-protein insertion into the ER membrane. Reason: Correct; GET3 associates with the ER membrane during TA delivery. Supporting Evidence: PMID:31461301 mediates insertion of TA (tail-anchored) proteins into the endoplasmic reticulum (ER) membrane
GO:0016887 ATP hydrolysis activity	IDA PMID:8884272 Isolation of the ATP-binding human homolog of the arsA compo...	ACCEPT	Summary: Direct biochemical demonstration that the human ArsA homolog (hARSA-I/ASNA1) is an ATPase. This is the experimental basis for GET3's core ATPase MF. Reason: Core molecular function with direct biochemical (IDA) support. Supporting Evidence: PMID:8884272 hARSA-I is an ATPase
GO:0016887 ATP hydrolysis activity	IDA PMID:9712828 Biochemical characterization of the human arsenite-stimulate...	ACCEPT	Summary: Direct biochemical characterization of the recombinant human protein measuring basal ATPase activity and ATP kinetics. Core ATPase MF. Reason: Core molecular function with direct biochemical (IDA) support; KM/Vmax for ATP determined. Supporting Evidence: PMID:9712828 ATPase activity
GO:0005789 endoplasmic reticulum membrane	NAS PMID:32910895 Structural Basis of Tail-Anchored Membrane Protein Biogenesi...	ACCEPT	Summary: ComplexPortal NAS assertion of ER membrane localization, reflecting the GET complex (within which GET3 docks) residing at the ER membrane. Reason: Correct; GET3 associates with the ER membrane GET complex during TA handoff. Supporting Evidence: PMID:32910895 an insertase (yeast Get1/Get2 or mammalian WRB/CAML) that captures the TA from a cytoplasmic chaperone
GO:0043529 GET complex	IPI PMID:32910895 Structural Basis of Tail-Anchored Membrane Protein Biogenesi...	ACCEPT	Summary: ComplexPortal IPI assignment of GET complex membership from the cryo-EM structure of the human GET insertase complex with bound GET3/TRC40. Reason: Core cellular component; structurally demonstrated. Supporting Evidence: PMID:32910895 captures the TA from a cytoplasmic chaperone (Get3 or TRC40, respectively)
GO:0045048 protein insertion into ER membrane	NAS PMID:23041287 Molecular machinery for insertion of tail-anchored membrane ...	KEEP AS NON CORE	Summary: ComplexPortal NAS assertion of the (parent) ER protein-insertion process; GET3/TRC40 targets TA proteins to the WRB/CAML receptor for insertion. Reason: Correct parent of GO:0071816; redundant general term. Supporting Evidence: PMID:23041287 an ATPase targeting newly synthesized TA proteins
GO:0071816 tail-anchored membrane protein insertion into ER membrane	IMP PMID:31461301 Biallelic Variants in ASNA1, Encoding a Cytosolic Targeting ...	ACCEPT	Summary: Mutant-phenotype evidence that the disease-associated Val163Ala ASNA1 mutant, while still able to capture a TA substrate, is inefficient in facilitating TA insertion into the ER membrane; asna1-null zebrafish show cardiac failure. Establishes GET3's role in the TA-insertion pathway. Reason: Core biological process with mutant-phenotype (IMP) support directly linking ASNA1 function to TA insertion. Supporting Evidence: PMID:31461301 inefficient in facilitating TA protein insertion into the ER membrane
GO:0140597 protein carrier activity	IDA PMID:23610396 Precise timing of ATPase activation drives targeting of tail...	ACCEPT	Summary: GET3/TRC40 is the TMD chaperone/carrier that shields the hydrophobic TA transmembrane domain and delivers it to the ER, harnessing ATP to drive TA membrane localization. This protein-carrier (chaperone) activity is a core molecular function complementary to its ATPase activity. Reason: Core molecular function; GET3 carries the TA-protein cargo as a soluble TMD chaperone, demonstrated by IDA. Supporting Evidence: PMID:23610396 Get3 harnesses the energy from ATP to drive PMID:37963916 the Get3 chaperone captures the TA protein substrate and delivers it to the Get1/Get2 membrane protein complex (GET insertase)
GO:0043529 GET complex	IDA PMID:32910895 Structural Basis of Tail-Anchored Membrane Protein Biogenesi...	ACCEPT	Summary: Direct structural evidence (cryo-EM) placing GET3/TRC40 within the GET insertase complex. Reason: Core cellular component; structurally demonstrated. Supporting Evidence: PMID:32910895 captures the TA from a cytoplasmic chaperone (Get3 or TRC40, respectively)
GO:0071816 tail-anchored membrane protein insertion into ER membrane	IMP PMID:23041287 Molecular machinery for insertion of tail-anchored membrane ...	ACCEPT	Summary: Mutant-phenotype evidence supporting GET3/TRC40's role as the ATPase targeting TA proteins for insertion, delivered to the CAML/WRB receptor complex. Reason: Core biological process with IMP support. Supporting Evidence: PMID:23041287 an ATPase targeting newly synthesized TA proteins
GO:0043529 GET complex	IPI PMID:23041287 Molecular machinery for insertion of tail-anchored membrane ...	ACCEPT	Summary: IPI identification of GET3/TRC40 in the receptor (GET) complex with WRB and CAML. Reason: Core cellular component; demonstrated by complex identification. Supporting Evidence: PMID:23041287 CAML and WRB as components of the TRC40 receptor complex
GO:0071816 tail-anchored membrane protein insertion into ER membrane	IDA PMID:25535373 Bag6 complex contains a minimal tail-anchor-targeting module...	ACCEPT	Summary: Direct evidence that the minimal Bag6 complex facilitates TA substrate transfer from SGTA to TRC40/GET3, the loading step that commits TA proteins to the GET targeting pathway. Reason: Core biological process; IDA demonstrating the TA-loading step onto GET3. Supporting Evidence: PMID:25535373 substrate transfer from small glutamine-rich tetratricopeptide repeat-containing
GO:0070062 extracellular exosome	HDA PMID:19056867 Large-scale proteomics and phosphoproteomics of urinary exos...	KEEP AS NON CORE	Summary: High-throughput urinary exosome proteomics catalog hit. Not indicative of a core localization or function for GET3. Reason: Proteomic catalog localization; not part of GET3's core cytosolic targeting function. Supporting Evidence: PMID:19056867 Large-scale proteomics and phosphoproteomics of urinary exosomes
GO:0005730 nucleolus	TAS PMID:9736449 Dual cytoplasmic and nuclear distribution of the novel arsen...	KEEP AS NON CORE	Summary: Legacy nucleolar localization reported in the early arsenite-ATPase characterization (cytoplasmic, perinuclear, and nucleolar distribution). Secondary to the core cytosolic targeting function. Reason: Legacy localization from early arsA-homolog work; not part of the core GET-pathway function. Supporting Evidence: PMID:9736449 Dual cytoplasmic and nuclear distribution of the novel arsenite-stimulated human ATPase
GO:0005737 cytoplasm	TAS PMID:9736449 Dual cytoplasmic and nuclear distribution of the novel arsen...	ACCEPT	Summary: Cytoplasmic localization from the early arsenite-ATPase study, consistent with GET3's primary cytosolic site of action. Reason: Correct primary compartment; corroborated by experimental EXP evidence. Supporting Evidence: PMID:9736449 Dual cytoplasmic and nuclear distribution

Core Functions

Cytosolic ATPase and TMD chaperone of the GET/TRC pathway that recognizes and binds the C-terminal transmembrane domain of newly synthesized tail-anchored proteins and, in an ATP-driven cycle, carries and delivers them to the ER membrane receptor for insertion.

Molecular Function:

ATP hydrolysis activity

Directly Involved In:

tail-anchored membrane protein insertion into ER membrane

Cellular Locations:

cytoplasm

Supporting Evidence:

PMID:17382883
cytosolic TMD recognition complex (TRC) that targets TA proteins for insertion into the ER membrane
file:human/GET3/GET3-uniprot.txt
ATP hydrolysis is required for insertion.

Soluble protein carrier (TMD chaperone) that shields the hydrophobic tail anchor of TA substrates and, as part of the GET complex docking on the WRB/CAML insertase, delivers them for ER membrane insertion.

Molecular Function:

protein carrier activity

Directly Involved In:

tail-anchored membrane protein insertion into ER membrane

In Complex:

GET complex

Supporting Evidence:

PMID:23610396
Get3 harnesses the energy from ATP to drive
file:human/GET3/GET3-uniprot.txt
Recognizes and selectively binds the transmembrane domain of TA

References

GO_REF:0000033

Annotation inferences using phylogenetic trees

GO_REF:0000044

Gene Ontology annotation based on UniProtKB/Swiss-Prot Subcellular Location vocabulary mapping

GO_REF:0000052

Gene Ontology annotation based on curation of immunofluorescence data

GO_REF:0000104

Electronic Gene Ontology annotations created by transferring manual GO annotations between related proteins based on shared sequence features

GO_REF:0000120

Combined Automated Annotation using Multiple IEA Methods

PMID:17382883

Identification of a targeting factor for posttranslational membrane protein insertion into the ER.

Identified the cytosolic TMD recognition complex (TRC); the 40 kDa ATPase subunit TRC40 is Asna-1, which targets TA proteins for ER insertion with release dependent on ATP hydrolysis.

PMID:19056867

Large-scale proteomics and phosphoproteomics of urinary exosomes.

PMID:21444755

WRB is the receptor for TRC40/Asna1-mediated insertion of tail-anchored proteins into the ER membrane.

TRC40/Asna1 (Get3 in yeast) is a conserved cytosolic ATPase that recognizes the TMD of TA proteins and delivers them to the ER membrane receptor WRB.

PMID:21516116

Next-generation sequencing to generate interactome datasets.

PMID:21911467

Insight into bacterial virulence mechanisms against host immune response via the Yersinia pestis-human protein-protein interaction network.

PMID:23041287

Molecular machinery for insertion of tail-anchored membrane proteins into the endoplasmic reticulum membrane in mammalian cells.

CAML and WRB are components of the TRC40 receptor complex; TRC40 is an ATPase targeting newly synthesized TA proteins to the ER for insertion.

PMID:23610396

Precise timing of ATPase activation drives targeting of tail-anchored proteins.

Get3 coordinates delivery of TA proteins to the ER; the Get4/5 loading complex locks Get3 in the ATP-bound state while the TA substrate activates Get3's ATPase ~100-fold, and Get3 harnesses ATP energy to drive TA membrane localization.

PMID:25416956

A proteome-scale map of the human interactome network.

PMID:25535373

Bag6 complex contains a minimal tail-anchor-targeting module and a mock BAG domain.

The minimal Bag6 complex facilitates TA substrate transfer from SGTA to TRC40, the loading step of the GET pathway.

PMID:28514442

Architecture of the human interactome defines protein communities and disease networks.

PMID:31461301

Biallelic Variants in ASNA1, Encoding a Cytosolic Targeting Factor of Tail-Anchored Proteins, Cause Rapidly Progressive Pediatric Cardiomyopathy.

ASNA1 (TRC40/GET3) is a ubiquitously expressed cytosolic chaperone mediating TA-protein insertion into the ER; the disease Val163Ala mutant captures TA but is inefficient in facilitating insertion, and asna1-null zebrafish develop cardiac failure.

PMID:32296183

A reference map of the human binary protein interactome.

PMID:32910895

Structural Basis of Tail-Anchored Membrane Protein Biogenesis by the GET Insertase Complex.

Cryo-EM of the GET insertase complex; the WRB/CAML (Get1/Get2) insertase captures the TA from the cytoplasmic chaperone GET3/TRC40 that GET3 binds as a homodimer.

PMID:33961781

Dual proteome-scale networks reveal cell-specific remodeling of the human interactome.

PMID:40205054

Multimodal cell maps as a foundation for structural and functional genomics.

PMID:8884272

Isolation of the ATP-binding human homolog of the arsA component of the bacterial arsenite transporter.

Isolated the human homolog of bacterial arsA (hARSA-I/ASNA1) and showed it is an ATPase analogous to bacterial ArsA.

PMID:9712828

Biochemical characterization of the human arsenite-stimulated ATPase (hASNA-I).

Recombinant hASNA-I exhibits basal ATPase activity (modestly arsenite-stimulated; antimonite does not stimulate), with measured ATP kinetics.

PMID:9736449

Dual cytoplasmic and nuclear distribution of the novel arsenite-stimulated human ATPase (hASNA-I).

Early study reporting a cytoplasmic, perinuclear, and nucleolar distribution of the arsenite-stimulated ATPase.

PMID:34264263

Capture and delivery of tail-anchored proteins to the endoplasmic reticulum.

Comprehensive review of the GET/TRC pathway; GET3/TRC40 is the central cytosolic homodimeric ATPase that captures TA substrates, shields the hydrophobic TMD, and delivers them to the ER WRB/CAML receptor-insertase for insertion.

PMID:36640319

The Get1/2 insertase forms a channel to mediate the insertion of tail-anchored proteins into the ER.

Get3 (TRC40) delivers TA proteins to the Get1/2 channel; Get3 binding seals the dynamically opening Get1/2 channel, and channel activity is required to release TA proteins from Get3 for insertion, defining the GET3-to-insertase handoff step.

PMID:37963916

The GET insertase exhibits conformational plasticity and induces membrane thinning.

Structures and simulations of human and C. thermophilum Get1/Get2/Get3 show that the gating interaction between Get2 helix alpha3' and Get3 drives conformational changes in both Get3 and the Get1/Get2 heterotetramer, promoting Get3 opening, nucleotide release, and TA substrate transfer to the membrane.

file:human/GET3/GET3-uniprot.txt

UniProt entry O43681 (GET3_HUMAN), ATPase ASNA1/TRC40

ATPase required for post-translational delivery of TA proteins to the ER; recognizes the TA TMD in the cytosol, targets to the GET1/WRB-CAMLG/GET2 receptor where ATP hydrolysis drives release/insertion; component of the GET complex; cytoplasmic/ER/nucleolar.

Suggested Experiments

Experiment: Reconstitute the full handoff cascade (SGTA -> BAG6/UBL4A/GET4 -> GET3 -> WRB/CAML) with purified components to quantify how GET4 priming and substrate-induced ATPase activation set the timing of TA capture and release.

Experiment: Define the endogenous GET3/ASNA1 TA-substrate repertoire in cardiomyocytes by proximity labeling and compare wild-type versus the Val163Ala disease variant to identify the TA clients whose mislocalization drives cardiomyopathy.

Deep Research

Falcon

(GET3-deep-research-falcon.md)

Research report: Human GET3 / ASNA1 / TRC40 (UniProt O43681) Falcon Edison Scientific Literature 21 citations 2 artifacts 2026-06-12T02:08:55.358391

Edison artifact artifact-00 (text/markdown)

## Context ID: pqac-00000016 The requested schematic figures of the GET insertase and its structural features are found in Figure 1 and Figure 6. Figure 1 provi (image/png)

The research report should be a detailed narrative explaining the function, biological processes, and localization of the gene product. Citations should be given for all claims.

You should prioritize authoritative reviews and primary scientific literature when conducting research. You can supplement
this with annotations you find in gene/protein databases, but these can be outdated or inaccurate.

We are specifically interested in the primary function of the gene - for enzymes, what reaction is catalyzed, and what is the substrate specificity? For transporters, what is the substrate? For structural proteins or adapters, what is the broader structural role? For signaling molecules, what is the role in the pathway.

We are interested in where in or outside the cell the gene product carries out its function.

We are also interested in the signaling or biochemical pathways in which the gene functions. We are less interested in broad pleiotropic effects, except where these elucidate the precise role.

Include evidence where possible. We are interested in both experimental evidence as well as inference from structure, evolution, or bioinformatic analysis. Precise studies should be prioritized over high-throughput, where available.

Research report: Human GET3 / ASNA1 / TRC40 (UniProt O43681)

0) Target verification (gene/protein identity)

UniProt accession O43681 corresponds to the human protein widely referred to as ASNA1 and TRC40, and explicitly described in the TA-protein targeting literature as the mammalian ortholog of yeast Get3 (hence the UniProt “GET3” name). The retrieved reviews and primary studies consistently use the mapping Get3 (yeast) ↔ TRC40/ASNA1 (metazoans/humans) in the context of the GET/TRC pathway for tail-anchored (TA) membrane protein insertion into the ER. (farkas2021captureanddelivery pages 1-3, mcdowell2023thegetinsertase pages 1-2, qin2023targetingandsurveillance pages 1-2)

1) Key concepts and definitions (current understanding)

Tail-anchored (TA) proteins

TA proteins are single-pass membrane proteins defined by a single C-terminal transmembrane domain (TMD) that anchors them in organelle membranes, leaving most of the protein cytosolic and a short C-terminal tail in the lumen (ER or other organelles). (farkas2021captureanddelivery pages 1-3, qin2023targetingandsurveillance pages 1-2)

GET/TRC pathway (ER targeting route for TA proteins)

The GET pathway in yeast and the conserved mammalian TRC (transmembrane recognition complex) pathway are major post-translational routes for targeting and inserting relatively hydrophobic TA proteins into the endoplasmic reticulum (ER) membrane. (farkas2021captureanddelivery pages 1-3, qin2023targetingandsurveillance pages 1-2)

ASNA1/TRC40/GET3 (central ATPase/chaperone)

Human ASNA1/TRC40 (GET3) is the central cytosolic targeting factor in this pathway: it is a homodimeric ATPase that binds TA clients, shields the hydrophobic TMD in a protected groove/pocket, and delivers the client to an ER membrane receptor/insertase for insertion. (farkas2021captureanddelivery pages 1-3, mcdowell2023thegetinsertase pages 1-2)

2) Molecular function and biochemical mechanism

2.1 Enzymatic activity

ASNA1/TRC40 is a P-loop NTPase ATPase whose functional cycle is coupled to TA-client handling (capture, delivery, release/recycling). In the conserved model described for Get3/TRC40, nucleotide state governs large conformational changes that regulate substrate binding and release. (farkas2021captureanddelivery pages 1-3, najdrova2022conservedmechanismfor pages 63-66)

2.2 Substrate specificity (what does ASNA1/TRC40 act on?)

The primary substrate class is tail-anchored membrane proteins, i.e., proteins whose key targeting determinant is a hydrophobic C-terminal TMD that must be protected from aggregation in the cytosol and then inserted into the ER membrane. (farkas2021captureanddelivery pages 1-3, mcdowell2023thegetinsertase pages 1-2, qin2023targetingandsurveillance pages 1-2)

2.3 Stepwise pathway model (from synthesis to ER insertion)

A consensus pathway model (reviews + recent mechanistic work) is:

Pre-targeting capture (cytosol): nascent TA proteins are initially captured by a pre-targeting complex (in mammals, SGTA and the BAG6 complex involving TRC35/UBL4A/BAG6) to prevent aggregation and to stage handoff. (farkas2021captureanddelivery pages 1-3, qin2023targetingandsurveillance pages 1-2)
Handoff to ATP-bound TRC40/ASNA1: TA substrates are transferred to ATP-bound TRC40, which shields the TA TMD in a hydrophobic binding site. (farkas2021captureanddelivery pages 1-3)
Delivery to ER receptor/insertase: TRC40 delivers cargo to the ER receptor/insertase complex (WRB/CAML in mammals; Get1/Get2 in yeast terminology). Receptor engagement drives TRC40 rearrangements enabling insertion and recycling. (farkas2021captureanddelivery pages 1-3, qin2023targetingandsurveillance pages 1-2)

Mechanistic detail from 2023 structural work

McDowell et al. (2023) describe a mechanistic sequence in which the Get2 cytosolic domain initially engages the Get3/TRC40–TA complex, then Get1 coiled-coil engagement promotes opening of Get3/TRC40 and nucleotide release, facilitating transfer of the TA substrate to the membrane insertase environment. (mcdowell2023thegetinsertase pages 1-2)

3) Cellular localization (where does ASNA1/TRC40 function?)

ASNA1/TRC40 functions primarily in the cytosol as a targeting factor/chaperone, delivering TA clients to the ER membrane receptor/insertase (WRB/CAML), where insertion occurs. (farkas2021captureanddelivery pages 1-3, qin2023targetingandsurveillance pages 1-2)

4) Recent developments (prioritizing 2023–2024)

4.1 2023: GET insertase structural plasticity and membrane remodeling

McDowell et al. (Nature Communications, 2023-11-; https://doi.org/10.1038/s41467-023-42867-2) provide structures, simulations, and functional data for human Get1/Get2/Get3 complexes, arguing that the GET insertase is conformationally plastic and induces local membrane thinning near a hydrophilic groove to facilitate insertion. They also summarize that TA proteins constitute a sizable fraction of membrane proteins (~5% of eukaryotic membrane proteins in their framing). (mcdowell2023thegetinsertase pages 1-2)

Visual evidence from this study illustrates the insertase topology and the location of the hydrophilic groove and membrane thinning features. (mcdowell2023thegetinsertase media dee660dd, mcdowell2023thegetinsertase media 0a7e56a9)

4.2 2023: A channel model for Get1/2-mediated insertion

Heo et al. (Cell Reports, 2023-01-; https://doi.org/10.1016/j.celrep.2022.111921) propose and support a model in which the Get1/2 insertase forms a transient aqueous channel in reconstituted membranes to mediate TA insertion.

Key quantitative/mechanistic points reported include:
- An estimated channel diameter of ~2.5 nm (interpreted as requiring two Get1/2 complexes). (heo2023theget12insertase pages 1-3)
- The channel model is framed as helping overcome two energetic barriers: movement of a hydrophobic TMD across headgroups and translocation of a C-terminal hydrophilic tail (up to ~30 residues) across the membrane interior. (heo2023theget12insertase pages 1-3)
- Experimental modulation indicates Get3 can affect channel behavior (e.g., reduced channel-series frequency and occupancy in microfluidics assays when Get3 is present vs BSA). (heo2023theget12insertase pages 29-34)

4.3 2023: Updated reviews of TA targeting and surveillance

Qin et al. (The Innovation Life, 2023-01-; https://doi.org/10.59717/j.xinn-life.2023.100013) emphasize the conserved pathway architecture and articulate a chaperone cascade view of ordered handoffs (Hsp70 → SGTA/Sgt2 → TRC components → TRC40/ASNA1 → WRB/CAML). This review also highlights that mammals add BAG6 as a bridging component in the pre-targeting complex. (qin2023targetingandsurveillance pages 1-2)

4.4 2024: Redox-regulated functional switching (TRC40 as a stress chaperone)

Dempsey et al. (bioRxiv preprint, 2024-07-10; https://doi.org/10.1101/2024.07.10.602939) report a redox-regulated switch in TRC40/ASNA1 function: besides its established ATP-dependent TA-targeting role, oxidative conditions (and/or ATP depletion) can convert TRC40 into an oligomeric chaperone that limits protein aggregation during stress.

Key quantitative/mechanistic points include:
- Oxidative activation tested with 2 mM H2O2/CuCl2 (or thiol-alkylation by NEM) promoted chaperone activity and inactivated ATPase-linked TA targeting behavior, reversible with reducing conditions and Mg-ATP. (dempsey2024adynamicredox pages 4-8)
- Reduced TRC40 is dimeric, whereas oxidized/thiol-blocked TRC40 forms tetramers and higher oligomers; tetramers are described as the smallest chaperone-active unit. (dempsey2024adynamicredox pages 4-8)
- Nucleotide binding can prevent oxidative activation, whereas ATP depletion facilitates oxidation-driven oligomerization and chaperone behavior; in cells, TRC40 forms reversible stress foci overlapping with Hsp70/Hsp110 and contributes to limiting stress-induced aggregation. (dempsey2024adynamicredox pages 4-8)

5) Current applications and real-world implementations

5.1 Functional genomics and proteomics “client spectrum” mapping at the ER

Jung & Zimmermann (International Journal of Molecular Sciences, 2023-09-; https://doi.org/10.3390/ijms241814166) summarize how systematic perturbation (siRNA/CRISPR) combined with quantitative proteomics can be used to infer client spectra of membrane targeting/insertion routes at the human ER, including TRC/GET components (TRC35/TRC40) versus SRP/SND/EMC/Sec61-related components. This is an applied, scalable approach to map which membrane-protein features route them into TRC/GET versus other pathways. (qin2023targetingandsurveillance pages 1-2)

5.2 Stress biology / proteostasis (emerging)

The 2024 redox-switch work positions TRC40/ASNA1 as a conditionally activated anti-aggregation factor under ATP-depleting oxidative stress, conceptually extending its “hydrophobic segment handling” function from TA biogenesis to broader proteostasis. While preclinical, this suggests an application space in stress-response biology and potentially in designing interventions that modulate redox/ATP states to influence TRC40 functional modes. (dempsey2024adynamicredox pages 4-8)

5.3 Quality control implications for TA biogenesis

Proteotoxic stress can disrupt the pre-targeting machinery upstream of TRC40. For example, Hagiwara et al. (Biochemical Journal, 2023-10-; https://doi.org/10.1042/bcj20230267) report that proteotoxic stresses can stimulate dissociation within the BAG6-associated complex (BAG6–UBL4A), implying that pathological aggregation stress may compromise efficient TA-protein biogenesis. This supports a real-world relevance to neurodegeneration-like proteotoxic conditions, although direct clinical translation remains indirect at present. (qin2023targetingandsurveillance pages 1-2)

6) Expert opinions and analysis (authoritative interpretation)

Consensus view of TRC40/ASNA1 as a specialized ATPase chaperone for C-terminal hydrophobes: Recent reviews and primary work agree that the TRC40/GET3 system is specialized for handling and delivering late-emerging hydrophobic segments (C-terminal TMDs) that cannot use canonical co-translational SRP targeting. (farkas2021captureanddelivery pages 1-3, qin2023targetingandsurveillance pages 1-2)
Mechanistic convergence on “protected hydrophobe handling + dedicated insertase”: 2023 mechanistic papers converge on the idea that successful insertion relies on both (i) cytosolic shielding by TRC40/Get3 and (ii) a dedicated membrane insertase (Get1/2; WRB/CAML context in mammals) that provides a specialized environment (hydrophilic groove/channel-like behavior) and can remodel the local bilayer to reduce insertion barriers. (heo2023theget12insertase pages 1-3, mcdowell2023thegetinsertase pages 1-2, mcdowell2023thegetinsertase media dee660dd)
Functional plasticity under stress as an emerging theme: The 2024 redox-switch preprint proposes that TRC40 may act as a “dual-mode” protein—TA targeting under normal energy/redox conditions, and oligomeric chaperoning under ATP-depleting oxidative stress—suggesting a broader conceptual role in cytosolic management of hydrophobic/aggregation-prone intermediates. This is a new hypothesis with evidence but awaits further peer-reviewed consolidation. (dempsey2024adynamicredox pages 4-8)

7) Relevant statistics and quantitative data points (from recent studies)

Prevalence: TA proteins are described as roughly ~5% of eukaryotic membrane proteins (McDowell et al., 2023). (mcdowell2023thegetinsertase pages 1-2)
Insertase physical model: A Get1/2 insertase channel diameter estimated at ~2.5 nm (Heo et al., 2023). (heo2023theget12insertase pages 1-3)
Client-tail constraint: The insertion problem includes translocation of C-terminal hydrophilic tails up to ~30 residues (Heo et al., 2023). (heo2023theget12insertase pages 1-3)
Stress/redox activation conditions: 2 mM H2O2/CuCl2 used to promote TRC40 oxidative activation; redox-linked oligomerization (dimer → tetramers/higher oligomers) reported as functionally important for chaperone activity (Dempsey et al., 2024). (dempsey2024adynamicredox pages 4-8)

8) Notes on “arsenite-stimulated ATPase” nomenclature

Some older classifications and cross-species annotations describe Get3/ASNA1 family members as “arsenite-stimulated ATPases” or “arsenical pump-driving ATPases.” In the retrieved corpus, recent mechanistic emphasis for the human protein is on TA-protein targeting/insertion and redox-sensitive functional switching, while arsenite-related ATPase nomenclature persists mostly as historical/family context rather than as an established physiological “arsenic pump” role in human cells. (kumar2021aconservedguided pages 31-33, kumar2021aconservedguided pages 7-9, dempsey2024adynamicredox pages 4-8)

9) Summary for functional annotation (human UniProt O43681)

Human GET3/ASNA1/TRC40 (O43681) is best annotated as a cytosolic ATP-dependent targeting factor/chaperone that binds and shields tail-anchored protein C-terminal TMDs, delivering them to the ER WRB/CAML receptor/insertase system for membrane insertion (TRC pathway). In 2023, structural and functional studies strengthened a physical mechanism in which the ER insertase presents a hydrophilic groove/channel-like environment and can locally remodel membrane thickness to facilitate insertion. In 2024, a redox/ATP-dependent switch was proposed that converts TRC40 into an oligomeric stress chaperone that protects cells from protein aggregation under ATP-depleting oxidative conditions. (farkas2021captureanddelivery pages 1-3, heo2023theget12insertase pages 1-3, mcdowell2023thegetinsertase pages 1-2, dempsey2024adynamicredox pages 4-8)

Evidence synopsis table

Concept / paper	Main finding for human ASNA1/TRC40 (GET3)	Pathway components highlighted	Quantitative / statistical points in evidence	URL	Publication date	Citation
Core function overview	Human ASNA1/TRC40 is the metazoan Get3 ortholog: a homodimeric ATPase/chaperone that captures tail-anchored (TA) proteins post-translationally, shields their hydrophobic C-terminal transmembrane domain, and delivers them to the ER for insertion.	Cytosolic TRC40/ASNA1 plus ER receptor/insertase WRB-CAML; upstream pre-targeting factors hand cargo to ATP-bound TRC40.	TA proteins are estimated to comprise ~5% of eukaryotic membrane proteins in one 2023 structural study.	https://doi.org/10.1083/jcb.202105004 ; https://doi.org/10.1038/s41467-023-42867-2	2021-07 ; 2023-11	(farkas2021captureanddelivery pages 1-3, mcdowell2023thegetinsertase pages 1-2)
Pathway organization review (Qin 2023 The Innovation Life)	The conserved TRC/GET pathway routes relatively hydrophobic TA proteins to the ER; TRC40/ASNA1 is the central ATPase receiving substrates from the pre-targeting machinery and passing them to the membrane receptor.	SGTA/Sgt2, TRC35/Get4, UBL4A/Get5, BAG6 in mammals; receptor complex WRB and CAML at ER.	No specific numerical result in the excerpt; emphasis is on ordered substrate handoff.	https://doi.org/10.59717/j.xinn-life.2023.100013	2023-01	(qin2023targetingandsurveillance pages 1-2)
Structural plasticity of the GET insertase (McDowell 2023 Nat Commun)	Structures of human/thermophilic Get1/Get2/Get3 show conserved insertase architecture, conformational plasticity, and local membrane thinning near a hydrophilic groove that likely lowers the energetic barrier for TA insertion. Get2 first engages Get3/TA, then Get1 promotes Get3 opening, nucleotide release, and cargo transfer.	Get3/TRC40 with Get1/Get2 insertase (human WRB/CAML orthologous system discussed in pathway context).	~5% of eukaryotic membrane proteins are TA; ~15° coiled-coil rotation reported between variant structures; buried surface area ~190 Å² in one interface description.	https://doi.org/10.1038/s41467-023-42867-2	2023-11	(mcdowell2023thegetinsertase pages 1-2, mcdowell2023thegetinsertase media dee660dd, mcdowell2023thegetinsertase media 0a7e56a9)
Channel model for insertion (Heo 2023 Cell Reports)	Get1/2 acts as an insertase/translocase forming a transient aqueous channel in membranes; channel activity helps release the TA substrate from Get3 and enables insertion. ATP binding then displaces Get3 for recycling.	Get3 cargo carrier; Get2 recruits Get3-TA; Get1 coiled-coil promotes substrate release; Get4/5 helps prevent premature rebinding.	Estimated channel diameter ~2.5 nm; hydrophilic tails up to ~30 residues discussed; channel-series frequency decreased ~10-fold and occupancy ~4-fold with Get3 versus BSA; replicates typically 2–5 with mean ± SD and unpaired t tests.	https://doi.org/10.1016/j.celrep.2022.111921	2023-01	(heo2023theget12insertase pages 1-3, heo2023theget12insertase pages 29-34)
Human ER client-spectrum analysis (Jung & Zimmermann 2023 IJMS)	Proteomic depletion studies in human cells support that TRC/GET preferentially handles membrane protein precursors with central or C-terminal topogenic sequences, consistent with ASNA1/TRC40 specialization for TA-like or late-emerging hydrophobic segments.	Human TRC pathway components including TRC35 and TRC40, compared with SRP, SND, EMC and Sec61-related pathways.	No single number quoted in the excerpt, but study is based on quantitative mass spectrometry across knockdown/knockout conditions.	https://doi.org/10.3390/ijms241814166	2023-09	(qin2023targetingandsurveillance pages 1-2)
Redox switch and stress chaperone role (Dempsey 2024 bioRxiv)	Beyond TA insertion, TRC40 can switch under ATP-depleting oxidative stress into a chaperone that limits protein aggregation; oxidation or thiol alkylation suppresses ATPase-linked TA targeting behavior and promotes stress-protective oligomerization.	TRC40/ASNA1 itself; stress-associated overlap with Hsp70/Hsp110-containing foci in cells.	Oxidative activation tested with 2 mM H2O2/CuCl2; reduced TRC40 is dimeric, oxidized/thiol-blocked TRC40 forms tetramers and higher oligomers; tetramers are the smallest chaperone-active unit.	https://doi.org/10.1101/2024.07.10.602939	2024-07	(dempsey2024adynamicredox pages 4-8)
Proteotoxic stress on pre-targeting complex (Hagiwara 2023 Biochem J)	Proteotoxic stress disrupts BAG6-UBL4A complex integrity, implying that stress can compromise the mammalian pre-targeting machinery required for efficient TA-protein biogenesis upstream of TRC40.	BAG6 and UBL4A (mammalian Get5 homolog), upstream of TRC40/Get3.	No specific number quoted in the excerpt.	https://doi.org/10.1042/bcj20230267	2023-10	(qin2023targetingandsurveillance pages 1-2)
Redox/arsenite-related legacy function	ASNA1/TRC40 belongs to the ArsA/Get3 ATPase family and earlier work characterized human hASNA1 as an arsenite-stimulated ATPase; more recent literature emphasizes TA targeting and redox-sensitive switching rather than arsenical pumping in human cells.	ArsA/Get3 family context; human TRC pathway function integrated with ATPase activity.	No additional quantitative value in the excerpt beyond historical classification.	https://doi.org/10.1101/2021.05.03.442402 ; https://doi.org/10.1101/2024.07.10.602939	2021-05 ; 2024-07	(kumar2021aconservedguided pages 31-33, kumar2021aconservedguided pages 7-9, dempsey2024adynamicredox pages 4-8)

Table: This table condenses the main mechanistic, structural, and stress-response findings for human ASNA1/TRC40 (GET3), with emphasis on 2023-2024 advances. It also captures the pathway components and quantitative data points most useful for functional annotation.

References

(farkas2021captureanddelivery pages 1-3): Ákos Farkas and Katherine E. Bohnsack. Capture and delivery of tail-anchored proteins to the endoplasmic reticulum. The Journal of Cell Biology, Jul 2021. URL: https://doi.org/10.1083/jcb.202105004, doi:10.1083/jcb.202105004. This article has 49 citations.
(mcdowell2023thegetinsertase pages 1-2): Melanie A. McDowell, Michael Heimes, Giray Enkavi, Ákos Farkas, Daniel Saar, Klemens Wild, Blanche Schwappach, Ilpo Vattulainen, and Irmgard Sinning. The get insertase exhibits conformational plasticity and induces membrane thinning. Nature Communications, Nov 2023. URL: https://doi.org/10.1038/s41467-023-42867-2, doi:10.1038/s41467-023-42867-2. This article has 16 citations and is from a highest quality peer-reviewed journal.
(qin2023targetingandsurveillance pages 1-2): Qing Qin, Kang Shen, and Xiangming Wang. Targeting and surveillance mechanisms for tail-anchored proteins. The Innovation Life, 1:100013, Jan 2023. URL: https://doi.org/10.59717/j.xinn-life.2023.100013, doi:10.59717/j.xinn-life.2023.100013. This article has 2 citations.
(najdrova2022conservedmechanismfor pages 63-66): V Najdrová. Conserved mechanism for targeting of tail-anchored proteins in eukaryotes. Unknown journal, 2022.
(mcdowell2023thegetinsertase media dee660dd): Melanie A. McDowell, Michael Heimes, Giray Enkavi, Ákos Farkas, Daniel Saar, Klemens Wild, Blanche Schwappach, Ilpo Vattulainen, and Irmgard Sinning. The get insertase exhibits conformational plasticity and induces membrane thinning. Nature Communications, Nov 2023. URL: https://doi.org/10.1038/s41467-023-42867-2, doi:10.1038/s41467-023-42867-2. This article has 16 citations and is from a highest quality peer-reviewed journal.
(mcdowell2023thegetinsertase media 0a7e56a9): Melanie A. McDowell, Michael Heimes, Giray Enkavi, Ákos Farkas, Daniel Saar, Klemens Wild, Blanche Schwappach, Ilpo Vattulainen, and Irmgard Sinning. The get insertase exhibits conformational plasticity and induces membrane thinning. Nature Communications, Nov 2023. URL: https://doi.org/10.1038/s41467-023-42867-2, doi:10.1038/s41467-023-42867-2. This article has 16 citations and is from a highest quality peer-reviewed journal.
(heo2023theget12insertase pages 1-3): Paul Heo, Jacob A. Culver, Jennifer Miao, Frederic Pincet, and Malaiyalam Mariappan. The get1/2 insertase forms a channel to mediate the insertion of tail-anchored proteins into the er. Cell Reports, 42:111921, Jan 2023. URL: https://doi.org/10.1016/j.celrep.2022.111921, doi:10.1016/j.celrep.2022.111921. This article has 8 citations and is from a highest quality peer-reviewed journal.
(heo2023theget12insertase pages 29-34): Paul Heo, Jacob A. Culver, Jennifer Miao, Frederic Pincet, and Malaiyalam Mariappan. The get1/2 insertase forms a channel to mediate the insertion of tail-anchored proteins into the er. Cell Reports, 42:111921, Jan 2023. URL: https://doi.org/10.1016/j.celrep.2022.111921, doi:10.1016/j.celrep.2022.111921. This article has 8 citations and is from a highest quality peer-reviewed journal.
(dempsey2024adynamicredox pages 4-8): Bianca Dempsey, Risai Dubrall, Olivia Chan, Kim Jasmin Lapacz, Jan Riemer, Ursula Jakob, and Kathrin Ulrich. A dynamic redox switch turns trc40 into a chaperone protecting human cells against atp-depleting, oxidative stress. bioRxiv, Jul 2024. URL: https://doi.org/10.1101/2024.07.10.602939, doi:10.1101/2024.07.10.602939. This article has 0 citations.
(kumar2021aconservedguided pages 31-33): Tarkeshwar Kumar, Satarupa Maitra, Abdur Rahman, and Souvik Bhattacharjee. A conserved guided entry of tail-anchored pathway is involved in the trafficking of tail-anchored membrane proteins in plasmodium falciparum. bioRxiv, May 2021. URL: https://doi.org/10.1101/2021.05.03.442402, doi:10.1101/2021.05.03.442402. This article has 2 citations.
(kumar2021aconservedguided pages 7-9): Tarkeshwar Kumar, Satarupa Maitra, Abdur Rahman, and Souvik Bhattacharjee. A conserved guided entry of tail-anchored pathway is involved in the trafficking of tail-anchored membrane proteins in plasmodium falciparum. bioRxiv, May 2021. URL: https://doi.org/10.1101/2021.05.03.442402, doi:10.1101/2021.05.03.442402. This article has 2 citations.

Artifacts

Edison artifact artifact-00

Citations

farkas2021captureanddelivery pages 1-3
mcdowell2023thegetinsertase pages 1-2
qin2023targetingandsurveillance pages 1-2
dempsey2024adynamicredox pages 4-8
najdrova2022conservedmechanismfor pages 63-66
kumar2021aconservedguided pages 31-33
kumar2021aconservedguided pages 7-9
https://doi.org/10.1038/s41467-023-42867-2
https://doi.org/10.1016/j.celrep.2022.111921
https://doi.org/10.59717/j.xinn-life.2023.100013
https://doi.org/10.1101/2024.07.10.602939
https://doi.org/10.3390/ijms241814166
https://doi.org/10.1042/bcj20230267
https://doi.org/10.1083/jcb.202105004
https://doi.org/10.1101/2021.05.03.442402
https://doi.org/10.1083/jcb.202105004,
https://doi.org/10.1038/s41467-023-42867-2,
https://doi.org/10.59717/j.xinn-life.2023.100013,
https://doi.org/10.1016/j.celrep.2022.111921,
https://doi.org/10.1101/2024.07.10.602939,
https://doi.org/10.1101/2021.05.03.442402,

📚 Additional Documentation

Notes

(GET3-notes.md)

GET3 (ASNA1 / TRC40) — review notes

UniProt: O43681 (GET3_HUMAN). HGNC:752. Synonyms: ARSA (HGNC), ASNA1, TRC40.
348 aa, homodimer, arsA ATPase family (HAMAP MF_03112). EC=3.6.4.-.

Core function: cytosolic ATPase that TARGETS tail-anchored (TA) proteins to the ER

GET3/TRC40/ASNA1 is the central cytosolic ATPase of the GET/TRC pathway. It selectively
recognizes the C-terminal transmembrane domain (TMD) of TA proteins in the cytosol, captures
them (handed off from SGTA via the GET4/UBL4A/BAG6 "bridging"/pre-targeting complex), and
delivers them to the ER-resident WRB/CAML (GET1/GET2) receptor-insertase, where ATP hydrolysis
drives TA release and insertion. GET3 is the targeting factor / chaperone, NOT the insertase
(insertase = WRB/CAML = GET1/GET2).

[file:human/GET3/GET3-uniprot.txt "ATPase required for the post-translational delivery of tail-anchored (TA) proteins to the endoplasmic reticulum"]
[file:human/GET3/GET3-uniprot.txt "Recognizes and selectively binds the transmembrane domain of TA proteins in the cytosol."]
[file:human/GET3/GET3-uniprot.txt "This complex then targets to the endoplasmic reticulum by membrane-bound receptors GET1/WRB and CAMLG/GET2, where the tail-anchored protein is released for insertion."]
PMID:17382883
PMID:17382883
PMID:17382883 — supports MUTAGEN G46R abolishing ATPase + dominantly inhibiting pathway (UniProt FT).
PMID:21444755
PMID:21444755
PMID:23041287

ATPase / ATP binding (MF)

PMID:9712828 — IDA ATP hydrolysis. Also gives KM/Vmax for ATP (UniProt biophysicochemical).
PMID:8884272 — IDA ATPase.
ATP binding/hydrolysis drives the targeting cycle: [file:human/GET3/GET3-uniprot.txt "ATP binding drives the homodimer towards the closed dimer state, facilitating recognition of newly synthesized TA membrane proteins. ATP hydrolysis is required for insertion."]
ATPase cycle regulated by upstream/downstream factors: PMID:23610396

Protein carrier / chaperone activity (MF)

GO:0140597 "protein carrier chaperone" (label in DR line; GOA labels it "protein carrier activity").
GET3 is a TMD chaperone that shields the hydrophobic TA-TMD and carries it to the ER.
- PMID:23610396
- PMID:23610396
- PMID:34887561

GET complex / receptor (CC + targeting)

[file:human/GET3/GET3-uniprot.txt "Component of the Golgi to ER traffic (GET) complex, which is composed of GET1/WRB, CAMLG/GET2 and GET3/TRC40"]
WRB = receptor: PMID:21444755
CAML = receptor, with WRB: PMID:23041287
Cryo-EM of GET insertase complex: PMID:32910895 — note: the INSERTASE is WRB/CAML; GET3 is the chaperone.
ComplexPortal CPX-6464 = GET complex; GOA GET complex part_of annotations (IDA/IPI PMID:32910895, IPI PMID:23041287) are correct.

Pre-targeting / bridging complex interactions (IPI partners)

GET4 (TRC35): [file:human/GET3/GET3-uniprot.txt "Interacts with GET4 (PubMed:34887561)."]; GOA IPI partners include UniProtKB:Q7L5D6 (GET4). PMID:34887561
CAMLG (P49069): [file:human/GET3/GET3-uniprot.txt "GET3 shows a higher affinity for CAMLG than for GET1"]; IPI partner UniProtKB:P49069 (CAMLG) in PMID:28514442, 33961781, 40205054, 32296183.
Bag6 complex transfers TA from SGTA to TRC40: PMID:25535373 — basis for GO:0071816 IDA (MUTAGEN K86D reduces pathway).
IntAct binary-interactome IPI captures (PMID:21516116, 25416956, 28514442, 32296183, 33961781, 40205054) include many TA-protein-like ER/secretory partners (AGR2, GPX7/8, PGRMC1/2, FKBP7, PDIA6, RCN1, TMX1, etc.) consistent with TA-client capture, plus GET4/CAMLG. Bare "protein binding" is uninformative; keep non-core. PMID:21911467 is a Yersinia pestis–human PPI screen (xeno partner yscD Q56975) — incidental, non-core.

Subcellular location

Cytoplasm/cytosol (site of action): [file:human/GET3/GET3-uniprot.txt "SUBCELLULAR LOCATION: Cytoplasm"]; EXP PMID:17382883, 21444755; TAS PMID:9736449.
ER: [file:human/GET3/GET3-uniprot.txt "Endoplasmic reticulum"]; EXP PMID:17382883, 21444755, 31461301. GET3 visits the ER membrane to dock on WRB/CAML. ER membrane (GO:0005789) NAS from ComplexPortal is reasonable as the GET complex resides there.
Nucleus/nucleolus: [file:human/GET3/GET3-uniprot.txt "Nucleus, nucleolus"]; reported in early arsenite-ATPase papers PMID:9736449 and HPA IDA. This is a secondary/legacy localization not tied to the core TA-targeting function — keep as non-core.
Extracellular exosome (HDA, PMID:19056867, urinary exosome proteomics): high-throughput proteomic catalog hit; non-core.

Legacy / secondary role: arsenite-stimulated ATPase (arsA homolog)

ASNA1 was originally cloned as the human homolog of bacterial arsA (arsenite transporter ATPase
component). The arsenite stimulation is modest and antimonite does NOT stimulate. This is a
legacy biochemical observation reflecting the ancestral arsA ATPase fold; the gene's bona fide
cellular role is TA-protein targeting. Treat arsenite/antimonite transport framing as
secondary/legacy.
- PMID:8884272
- PMID:9712828
- No arsenite/antimonite transport GO annotation actually appears in the current GOA (only ATP hydrolysis IDA from these papers) — so there is nothing to MARK_AS_OVER_ANNOTATED here; the IDA ATP-hydrolysis annotations are accepted as the genuine MF.

Disease

CMD2H (dilated cardiomyopathy 2H), autosomal recessive, rapidly progressive infantile.
PMID:31461301
Val163Ala mutant: folded protein captures TA but is "inefficient in facilitating TA protein
insertion into the ER membrane" — directly supports GO:0071816 IMP. asna1-null zebrafish
cardiac failure rescued by WT but not mutant human ASNA1 mRNA.

Annotation decisions summary

ATP hydrolysis (IBA, IEA, IDA x2): ACCEPT (core MF). ATP binding (IEA): KEEP_AS_NON_CORE (structural, subsidiary to ATPase).
GO:0071816 TA insertion into ER (IBA/IEA/IMP x2/IDA): ACCEPT as core BP for the experimental/IBA ones; the BP label says "insertion" but is the standard GO term for the GET-pathway role of the targeting factor — accept.
GO:0045048 protein insertion into ER membrane (IEA InterPro2GO; NAS ComplexPortal): KEEP_AS_NON_CORE / parent-of-71816, redundant general term.
GO:0140597 protein carrier activity/chaperone (IDA PMID:23610396): ACCEPT (core MF — TMD chaperone/carrier).
GET complex part_of (IEA/IPI/IDA): ACCEPT.
protein binding (IPI, 7 refs): KEEP_AS_NON_CORE (bare term; informative partners captured in notes/complex annotations).
cytoplasm (IEA/EXP/TAS): ACCEPT. ER (IEA/EXP): ACCEPT. ER membrane (NAS): ACCEPT (GET complex resides there).
nucleolus (IEA/IDA/TAS), nucleoplasm (IDA): KEEP_AS_NON_CORE (legacy/HPA localization, not core function).
extracellular exosome (HDA): KEEP_AS_NON_CORE (proteomic catalog).

Falcon deep-research findings (incorporated 2026-06)

GET3/TRC40 chaperone-captures the TA substrate and delivers it to the Get1/Get2 (WRB/CAML) insertase, which inserts it via a membrane-embedded hydrophilic groove — confirms GET3 is the cytosolic carrier/chaperone, not the insertase. PMID:37963916. Added to supported_by for GO:0140597 protein carrier activity.
Receptor-driven conformational opening of GET3: the gating interaction between Get2 helix α3' and Get3 drives conformational changes in both Get3 and the Get1/Get2 heterotetramer, promoting Get3 opening and nucleotide release for substrate transfer. PMID:37963916. Mechanistic detail on the GET3 ATP/handoff cycle.
GET3 gates the insertase channel: in reconstituted membranes, Get3 (TRC40) binding seals the dynamically opening Get1/2 channel, and channel activity is required to release TA proteins from Get3 for insertion. PMID:36640319. Defines the GET3-to-insertase handoff step.
Pathway review (Farkas & Bohnsack 2021): GET3/TRC40 is the central cytosolic homodimeric ATPase that shields the TA TMD and delivers it to the ER WRB/CAML receptor-insertase; corroborates existing core-function calls. PMID:34264263 (added as HIGH-relevance reference).
Emerging (NOT added as structured reference — bioRxiv preprint, unverifiable on PubMed): Dempsey et al. 2024 [DOI:10.1101/2024.07.10.602939] propose a redox-regulated switch converting TRC40 from an ATP-dependent TA-targeting ATPase (reduced dimer) into an oligomeric anti-aggregation stress chaperone (oxidized/thiol-blocked tetramers+) under ATP-depleting oxidative stress, reversible with reducing conditions and Mg-ATP. Intriguing dual-mode hypothesis but preprint-only; treat as a candidate question/experiment, not an annotation.

Pn Notes

(GET3-pn-notes.md)

GET3 PN Consistency Notes

Generated: 2026-06-18
Project: PROTEOSTASIS
Scope: PN consistency rereview against local AIGR review and available deep-research artifacts
UniProt: O43681
AIGR review status: COMPLETE
Review batch: proteostasis-batch-2026-06-11
Batch change status: added

Source Files Checked

AIGR review YAML: present - genes/human/GET3/GET3-ai-review.yaml
AIGR review HTML: missing - genes/human/GET3/GET3-ai-review.html
Gene notes: present - genes/human/GET3/GET3-notes.md
GOA TSV: present - genes/human/GET3/GET3-goa.tsv
UniProt record: present - genes/human/GET3/GET3-uniprot.txt
PN dossier: projects/PROTEOSTASIS/reports/phase1_dossiers/GET3.md
PN consistency section: projects/PROTEOSTASIS/reports/phase1_dossiers/_sections/GET3.md
PN projection report: projects/PROTEOSTASIS/reports/pn_projection/pn_projected_gene_go_summary.tsv
PN mapping audit: projects/PROTEOSTASIS/reports/pn_mapping_audit/current_mapping_scrutiny.tsv

Deep Research Files

genes/human/GET3/GET3-deep-research-falcon.md

AIGR Review Snapshot

Description: GET3 (ASNA1; also known as TRC40 and arsenite-stimulated ATPase) is the central cytosolic ATPase of the GET/TRC pathway for post-translational targeting of tail-anchored (TA) membrane proteins to the endoplasmic reticulum (ER). It is a homodimeric P-loop NTPase of the ArsA/ArsA-like ATPase family. GET3 selectively recognizes and binds the single C-terminal transmembrane domain (TMD) of newly synthesized TA proteins in the cytosol, receiving them via the BAG6/UBL4A/GET4 pre-targeting and bridging machinery (and from the cochaperone SGTA), and shields the hydrophobic TMD as a soluble carrier/chaperone. ATP binding drives the homodimer into a closed state that captures the substrate; the GET3-TA complex then docks at the ER-membrane receptor-insertase formed by GET1/WRB and CAMLG/GET2, and ATP hydrolysis triggers TA release for insertion into the lipid bilayer, after which GET3 returns to the cytosol for another round. GET3 is thus the targeting factor and TMD chaperone of the pathway, not the membrane insertase itself. It was originally isolated as the human homolog of bacterial ArsA, an arsenite/antimonite-stimulated ATPase, but its physiological role is TA-protein biogenesis. Loss-of-function variants cause an autosomal recessive, rapidly progressive infantile dilated cardiomyopathy. GET3 acts predominantly in the cytoplasm and transiently at the ER membrane.
Existing/core annotation action counts: ACCEPT: 23; KEEP_AS_NON_CORE: 15

PN Consistency Summary

Consistency: Deep research, review YAML, and PN annotation are consistent: GET3/ASNA1/TRC40 is the cytosolic homodimeric ATPase + TMD chaperone (carrier) of the GET/TRC pathway — the targeting factor, NOT the insertase (insertase = WRB/CAML). The review cleanly distinguishes the ATPase MF (GO:0016887 ATP hydrolysis activity) and carrier/chaperone MF (GO:0140597 protein carrier activity) from the GET1 receptor role, exactly as the task brief requires. Legacy arsenite/nucleolar framing correctly treated as secondary/non-core.
PN story / NEW pressure: PN asserts post-translational ER targeting of TA proteins — already captured: GO:0071816 (involved_in), GO:0016887 + GO:0140597 (core MFs), GO:0043529 (GET complex), GO:0045048 (non-core parent). No NEW GO term warranted. GO:0006620 (post-translational protein targeting to ER membrane) is a defensible, verified-real term that fits GET3's targeting role well — arguably a good added BP for GET3, though the GET-pathway role is conventionally captured by GO:0071816 in GOA.
Evidence alignment: High overlap — review's targeting-pathway PMIDs (17382883, 21444755, 23041287, 23610396, 25535373, 31461301, 32910895, 36640319, 37963916) fully evidence the PN targeting claim.
Verdict: Consistent; exemplary review with correct ATPase-vs-chaperone MF split. GO:0006620 is an apt (verified) targeting term for GET3; flag the cross-gene goa_status inconsistency with the GET1 dossier.

Full Consistency Review

UniProt: O43681 (ASNA1/TRC40) · batch: proteostasis-batch-2026-06-11 · review status: COMPLETE
PN placement: ER proteostasis|Protein transport|GET pathway component ; PN-node mapping: group → GO:0006620 (post-translational protein targeting to ER membrane), scope=ok_for_propagation, goa_status=new_to_goa; class → GO:0015031 (protein transport); branch=no_mapping.
Consistency: Deep research, review YAML, and PN annotation are consistent: GET3/ASNA1/TRC40 is the cytosolic homodimeric ATPase + TMD chaperone (carrier) of the GET/TRC pathway — the targeting factor, NOT the insertase (insertase = WRB/CAML). The review cleanly distinguishes the ATPase MF (GO:0016887 ATP hydrolysis activity) and carrier/chaperone MF (GO:0140597 protein carrier activity) from the GET1 receptor role, exactly as the task brief requires. Legacy arsenite/nucleolar framing correctly treated as secondary/non-core.
PN story / NEW pressure: PN asserts post-translational ER targeting of TA proteins — already captured: GO:0071816 (involved_in), GO:0016887 + GO:0140597 (core MFs), GO:0043529 (GET complex), GO:0045048 (non-core parent). No NEW GO term warranted. GO:0006620 (post-translational protein targeting to ER membrane) is a defensible, verified-real term that fits GET3's targeting role well — arguably a good added BP for GET3, though the GET-pathway role is conventionally captured by GO:0071816 in GOA.
Mapping strategy: GET3 does not change the GET-node mapping. GO:0006620 fits GET3 (the targeting ATPase) more precisely than GET1 (the receptor-insertase). The goa_status new_to_goa here vs more_specific_than_existing_goa for the identical GET1 mapping is internally inconsistent across the two same-node genes — one needs correction.
Evidence alignment: High overlap — review's targeting-pathway PMIDs (17382883, 21444755, 23041287, 23610396, 25535373, 31461301, 32910895, 36640319, 37963916) fully evidence the PN targeting claim.
Verdict: Consistent; exemplary review with correct ATPase-vs-chaperone MF split. GO:0006620 is an apt (verified) targeting term for GET3; flag the cross-gene goa_status inconsistency with the GET1 dossier.

PN Dossier Context

review_batch: proteostasis-batch-2026-06-11
review_yaml: genes/human/GET3/GET3-ai-review.yaml
PN workbook rows: 1

PN row 1: ER proteostasis | Protein transport | GET pathway component

UniProt: O43681
In branches: ER
PN-node mapping records (path + ancestors):
- [group] ER proteostasis|Protein transport|GET pathway component
  status=mapped scope=ok_for_propagation_to_go GO=[GO:0006620 post-translational protein targeting to endoplasmic reticulum membrane]
  rationale: The PN GET-pathway group covers machinery for post-translational delivery of tail-anchored membrane proteins to the ER. GO does not model the GET pathway directly in the local cache, and the closest supported process term is post-translational targeting to the ER membrane.
- [class] ER proteostasis|Protein transport
  status=mapped scope=ok_for_propagation_to_go GO=[GO:0015031 protein transport]
  rationale: The PN ER Protein transport class groups ER-targeting and ER-insertion pathways. GO protein transport is the appropriate propagation target, while the source class remains ER-specific and broader than any single GO transport subtype.
- [branch] ER proteostasis
  status=no_mapping scope= GO=[]
  rationale: Reviewed as a top-level PN branch. This is a systems/taxonomy umbrella, not a direct GO assertion; narrower child curations carry any propagating GO mappings.

Projected GO annotations (2)

GO:0015031 protein transport | scope=ok_for_propagation_to_go | goa_status=new_to_goa | from=ER proteostasis|Protein transport
GO:0006620 post-translational protein targeting to endoplasmic reticulum membrane | scope=ok_for_propagation_to_go | goa_status=new_to_goa | from=ER proteostasis|Protein transport|GET pathway component

Note

This file is generated from the current PROTEOSTASIS phase-1 dossier and local gene-review artifacts. Edit the source review, PN mapping, or dossier rather than this generated note when correcting the underlying curation.

📄 View Raw YAML

id: O43681
gene_symbol: GET3
product_type: PROTEIN
status: COMPLETE
taxon:
  id: NCBITaxon:9606
  label: Homo sapiens
description: GET3 (ASNA1; also known as TRC40 and arsenite-stimulated ATPase) is the central cytosolic ATPase of the GET/TRC pathway for post-translational targeting of tail-anchored (TA) membrane proteins to the endoplasmic reticulum (ER). It is a homodimeric P-loop NTPase of the ArsA/ArsA-like ATPase family. GET3 selectively recognizes and binds the single C-terminal transmembrane domain (TMD) of newly synthesized TA proteins in the cytosol, receiving them via the BAG6/UBL4A/GET4 pre-targeting and bridging machinery (and from the cochaperone SGTA), and shields the hydrophobic TMD as a soluble carrier/chaperone. ATP binding drives the homodimer into a closed state that captures the substrate; the GET3-TA complex then docks at the ER-membrane receptor-insertase formed by GET1/WRB and CAMLG/GET2, and ATP hydrolysis triggers TA release for insertion into the lipid bilayer, after which GET3 returns to the cytosol for another round. GET3 is thus the targeting factor and TMD chaperone of the pathway, not the membrane insertase itself. It was originally isolated as the human homolog of bacterial ArsA, an arsenite/antimonite-stimulated ATPase, but its physiological role is TA-protein biogenesis. Loss-of-function variants cause an autosomal recessive, rapidly progressive infantile dilated cardiomyopathy. GET3 acts predominantly in the cytoplasm and transiently at the ER membrane.
existing_annotations:
- term:
    id: GO:0016887
    label: ATP hydrolysis activity
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: enables
  review:
    summary: Phylogenetic annotation of GET3's ATP hydrolysis activity, the catalytic core of its targeting cycle. Conserved across the ArsA/Get3 family.
    action: ACCEPT
    reason: Core molecular function; GET3 is an ATPase whose ATP hydrolysis drives TA-protein release/insertion, supported by IDA and EC 3.6.4.-.
    supported_by:
    - reference_id: file:human/GET3/GET3-uniprot.txt
      supporting_text: ATP hydrolysis is required for insertion.
- term:
    id: GO:0071816
    label: tail-anchored membrane protein insertion into ER membrane
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: involved_in
  review:
    summary: Phylogenetic annotation of GET3's defining biological process. This is the standard GO term used for the GET-pathway role; GET3 is the cytosolic targeting factor whose action commits TA proteins to ER insertion.
    action: ACCEPT
    reason: Core biological process; conserved and supported by experimental evidence (IMP/IDA).
    supported_by:
    - reference_id: file:human/GET3/GET3-uniprot.txt
      supporting_text: ATPase required for the post-translational delivery of tail-
- term:
    id: GO:0005524
    label: ATP binding
  evidence_type: IEA
  original_reference_id: GO_REF:0000120
  qualifier: enables
  review:
    summary: ATP binding drives the GET3 homodimer to the closed state that captures the TA substrate. A structural/mechanistic attribute subsidiary to the catalytic ATP hydrolysis activity.
    action: KEEP_AS_NON_CORE
    reason: Accurate (GET3 binds ATP) but subsidiary to the more informative ATP hydrolysis activity that represents the core MF.
    supported_by:
    - reference_id: file:human/GET3/GET3-uniprot.txt
      supporting_text: ATP binding drives the
- term:
    id: GO:0005730
    label: nucleolus
  evidence_type: IEA
  original_reference_id: GO_REF:0000044
  qualifier: located_in
  review:
    summary: Electronic transfer of a legacy nucleolar localization reported in early arsenite-ATPase studies. Not tied to the core cytosolic TA-targeting function.
    action: KEEP_AS_NON_CORE
    reason: Legacy/secondary localization derived from early arsA-homolog studies; not part of the core GET-pathway function.
    supported_by:
    - reference_id: file:human/GET3/GET3-uniprot.txt
      supporting_text: 'Nucleus, nucleolus'
- term:
    id: GO:0005737
    label: cytoplasm
  evidence_type: IEA
  original_reference_id: GO_REF:0000044
  qualifier: located_in
  review:
    summary: Electronic transfer of the cytoplasmic localization, the primary site where GET3 captures TA substrates. Consistent with experimental evidence.
    action: ACCEPT
    reason: Correct primary compartment; GET3 is a cytosolic targeting factor.
    supported_by:
    - reference_id: file:human/GET3/GET3-uniprot.txt
      supporting_text: 'SUBCELLULAR LOCATION: Cytoplasm'
- term:
    id: GO:0005783
    label: endoplasmic reticulum
  evidence_type: IEA
  original_reference_id: GO_REF:0000120
  qualifier: located_in
  review:
    summary: Electronic annotation of ER localization, where GET3 transiently docks on the WRB/CAML receptor to deliver TA substrates. Consistent with experimental evidence.
    action: ACCEPT
    reason: Correct; GET3 visits the ER membrane to hand off its TA cargo.
    supported_by:
    - reference_id: file:human/GET3/GET3-uniprot.txt
      supporting_text: 'Endoplasmic'
- term:
    id: GO:0016887
    label: ATP hydrolysis activity
  evidence_type: IEA
  original_reference_id: GO_REF:0000120
  qualifier: enables
  review:
    summary: Electronic assignment of the core ATP hydrolysis activity, consistent with experimental IDA and the catalytic activity record.
    action: ACCEPT
    reason: Correct core molecular function; redundant with IDA/IBA.
    supported_by:
    - reference_id: file:human/GET3/GET3-uniprot.txt
      supporting_text: ATP hydrolysis is required for insertion.
- term:
    id: GO:0045048
    label: protein insertion into ER membrane
  evidence_type: IEA
  original_reference_id: GO_REF:0000104
  qualifier: involved_in
  review:
    summary: Electronic (Swiss-Prot keyword/feature transfer) annotation of the parent ER protein-insertion process. Correct but less specific than the TA-insertion term.
    action: KEEP_AS_NON_CORE
    reason: Correct but generic parent of GO:0071816, which better captures GET3's role.
    supported_by:
    - reference_id: file:human/GET3/GET3-uniprot.txt
      supporting_text: ATPase required for the post-translational delivery of tail-
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:21516116
  qualifier: enables
  review:
    summary: High-throughput interactome screen capturing GET3 protein interactions. The bare protein binding term is uninformative.
    action: KEEP_AS_NON_CORE
    reason: Bare protein binding from a high-throughput screen; uninformative for the core MF.
    supported_by:
    - reference_id: PMID:21516116
      supporting_text: Next-generation sequencing to generate interactome datasets
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:21911467
  qualifier: enables
  review:
    summary: Yersinia pestis-human protein-protein interaction screen; the captured partner is a bacterial xenobiotic protein (yscD), an incidental cross-species interaction unrelated to GET3's function.
    action: KEEP_AS_NON_CORE
    reason: Incidental xenobiotic (bacterial) interaction from a host-pathogen screen; bare protein binding, not relevant to core function.
    supported_by:
    - reference_id: file:human/GET3/GET3-uniprot.txt
      supporting_text: 'O43681; Q56975: yscD'
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:25416956
  qualifier: enables
  review:
    summary: Proteome-scale human interactome map; source of IntAct partners including ER/secretory and TA-like proteins. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: High-throughput interactome; bare protein binding term is uninformative.
    supported_by:
    - reference_id: PMID:25416956
      supporting_text: A proteome-scale map of the human interactome network
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:28514442
  qualifier: enables
  review:
    summary: Human interactome architecture study; captures GET3 interactions including the functionally relevant CAMLG partner, but uses the uninformative bare protein binding term.
    action: KEEP_AS_NON_CORE
    reason: Records real interactions (including CAMLG) but bare protein binding is uninformative; functional partners are captured via GET complex membership.
    supported_by:
    - reference_id: file:human/GET3/GET3-uniprot.txt
      supporting_text: 'O43681; P49069: CAMLG'
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:32296183
  qualifier: enables
  review:
    summary: HuRI binary interactome (Y2H); source of many IntAct partners of GET3, including ER/secretory and TA-like proteins consistent with TA-client capture. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: High-throughput Y2H interactome; bare protein binding is uninformative.
    supported_by:
    - reference_id: PMID:32296183
      supporting_text: A reference map of the human binary protein interactome
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:33961781
  qualifier: enables
  review:
    summary: Proteome-scale cell-specific interactome network; captures GET3 interactions including CAMLG. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: High-throughput interactome; bare protein binding is uninformative.
    supported_by:
    - reference_id: file:human/GET3/GET3-uniprot.txt
      supporting_text: 'O43681; P49069: CAMLG'
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:40205054
  qualifier: enables
  review:
    summary: Multimodal cell map study; captures GET3 protein interactions including CAMLG. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: High-throughput interactome; bare protein binding is uninformative.
    supported_by:
    - reference_id: file:human/GET3/GET3-uniprot.txt
      supporting_text: 'O43681; P49069: CAMLG'
- term:
    id: GO:0043529
    label: GET complex
  evidence_type: IEA
  original_reference_id: GO_REF:0000120
  qualifier: part_of
  review:
    summary: Electronic assignment of GET complex membership. GET3/TRC40 is a defining subunit of the GET complex (GET1/WRB + CAMLG/GET2 + GET3/TRC40).
    action: ACCEPT
    reason: Core cellular component; consistent with experimental IDA/IPI evidence.
    supported_by:
    - reference_id: file:human/GET3/GET3-uniprot.txt
      supporting_text: Component of the Golgi to ER
- term:
    id: GO:0071816
    label: tail-anchored membrane protein insertion into ER membrane
  evidence_type: IEA
  original_reference_id: GO_REF:0000120
  qualifier: involved_in
  review:
    summary: Electronic assignment of the core TA-insertion process, consistent with experimental evidence.
    action: ACCEPT
    reason: Correct core process; redundant with IMP/IDA/IBA.
    supported_by:
    - reference_id: file:human/GET3/GET3-uniprot.txt
      supporting_text: ATPase required for the post-translational delivery of tail-
- term:
    id: GO:0005654
    label: nucleoplasm
  evidence_type: IDA
  original_reference_id: GO_REF:0000052
  qualifier: located_in
  review:
    summary: HPA immunofluorescence nucleoplasm localization. A secondary localization not connected to the core cytosolic TA-targeting function.
    action: KEEP_AS_NON_CORE
    reason: HPA-derived secondary localization; not part of the core GET-pathway function.
    supported_by:
    - reference_id: file:human/GET3/GET3-uniprot.txt
      supporting_text: 'Nucleus, nucleolus'
- term:
    id: GO:0005730
    label: nucleolus
  evidence_type: IDA
  original_reference_id: GO_REF:0000052
  qualifier: located_in
  review:
    summary: HPA immunofluorescence nucleolar localization, consistent with the legacy nucleolar distribution reported for the arsenite-ATPase. Secondary to the core function.
    action: KEEP_AS_NON_CORE
    reason: Secondary/legacy localization; not part of the core cytosolic targeting function.
    supported_by:
    - reference_id: file:human/GET3/GET3-uniprot.txt
      supporting_text: 'Nucleus, nucleolus'
- term:
    id: GO:0005737
    label: cytoplasm
  evidence_type: EXP
  original_reference_id: PMID:17382883
  qualifier: located_in
  review:
    summary: Direct experimental cytoplasmic localization from the study that identified TRC40/Asna-1 as the cytosolic TA-targeting ATPase.
    action: ACCEPT
    reason: Core compartment; GET3 acts as a cytosolic targeting factor.
    supported_by:
    - reference_id: PMID:17382883
      supporting_text: cytosolic TMD recognition complex (TRC) that targets TA proteins for insertion into the ER membrane
- term:
    id: GO:0005737
    label: cytoplasm
  evidence_type: EXP
  original_reference_id: PMID:21444755
  qualifier: located_in
  review:
    summary: Direct experimental cytoplasmic localization, consistent with GET3/TRC40 being a conserved cytosolic ATPase.
    action: ACCEPT
    reason: Core compartment; corroborated by multiple experimental sources.
    supported_by:
    - reference_id: PMID:21444755
      supporting_text: TRC40/Asna1 (Get3 in yeast)
- term:
    id: GO:0005783
    label: endoplasmic reticulum
  evidence_type: EXP
  original_reference_id: PMID:17382883
  qualifier: located_in
  review:
    summary: Direct experimental ER localization; GET3 docks on the ER membrane to deliver its TA cargo to the WRB/CAML receptor.
    action: ACCEPT
    reason: Correct; GET3 transiently associates with the ER during TA delivery.
    supported_by:
    - reference_id: PMID:17382883
      supporting_text: targets TA proteins for insertion into the ER membrane
- term:
    id: GO:0005783
    label: endoplasmic reticulum
  evidence_type: EXP
  original_reference_id: PMID:21444755
  qualifier: located_in
  review:
    summary: Direct experimental ER localization, consistent with GET3 delivering TA proteins to the ER membrane receptor.
    action: ACCEPT
    reason: Correct; GET3 visits the ER to hand off TA cargo.
    supported_by:
    - reference_id: PMID:21444755
      supporting_text: delivers them to the ER membrane for insertion
- term:
    id: GO:0005783
    label: endoplasmic reticulum
  evidence_type: EXP
  original_reference_id: PMID:31461301
  qualifier: located_in
  review:
    summary: Experimental ER localization from the ASNA1 cardiomyopathy study, in which ASNA1 mediates TA-protein insertion into the ER membrane.
    action: ACCEPT
    reason: Correct; GET3 associates with the ER membrane during TA delivery.
    supported_by:
    - reference_id: PMID:31461301
      supporting_text: mediates insertion of TA (tail-anchored) proteins into the endoplasmic reticulum (ER) membrane
- term:
    id: GO:0016887
    label: ATP hydrolysis activity
  evidence_type: IDA
  original_reference_id: PMID:8884272
  qualifier: enables
  review:
    summary: Direct biochemical demonstration that the human ArsA homolog (hARSA-I/ASNA1) is an ATPase. This is the experimental basis for GET3's core ATPase MF.
    action: ACCEPT
    reason: Core molecular function with direct biochemical (IDA) support.
    supported_by:
    - reference_id: PMID:8884272
      supporting_text: hARSA-I is an ATPase
- term:
    id: GO:0016887
    label: ATP hydrolysis activity
  evidence_type: IDA
  original_reference_id: PMID:9712828
  qualifier: enables
  review:
    summary: Direct biochemical characterization of the recombinant human protein measuring basal ATPase activity and ATP kinetics. Core ATPase MF.
    action: ACCEPT
    reason: Core molecular function with direct biochemical (IDA) support; KM/Vmax for ATP determined.
    supported_by:
    - reference_id: PMID:9712828
      supporting_text: ATPase activity
- term:
    id: GO:0005789
    label: endoplasmic reticulum membrane
  evidence_type: NAS
  original_reference_id: PMID:32910895
  qualifier: located_in
  review:
    summary: ComplexPortal NAS assertion of ER membrane localization, reflecting the GET complex (within which GET3 docks) residing at the ER membrane.
    action: ACCEPT
    reason: Correct; GET3 associates with the ER membrane GET complex during TA handoff.
    supported_by:
    - reference_id: PMID:32910895
      supporting_text: an insertase (yeast Get1/Get2 or mammalian WRB/CAML) that captures the TA from a cytoplasmic chaperone
- term:
    id: GO:0043529
    label: GET complex
  evidence_type: IPI
  original_reference_id: PMID:32910895
  qualifier: part_of
  review:
    summary: ComplexPortal IPI assignment of GET complex membership from the cryo-EM structure of the human GET insertase complex with bound GET3/TRC40.
    action: ACCEPT
    reason: Core cellular component; structurally demonstrated.
    supported_by:
    - reference_id: PMID:32910895
      supporting_text: captures the TA from a cytoplasmic chaperone (Get3 or TRC40, respectively)
- term:
    id: GO:0045048
    label: protein insertion into ER membrane
  evidence_type: NAS
  original_reference_id: PMID:23041287
  qualifier: involved_in
  review:
    summary: ComplexPortal NAS assertion of the (parent) ER protein-insertion process; GET3/TRC40 targets TA proteins to the WRB/CAML receptor for insertion.
    action: KEEP_AS_NON_CORE
    reason: Correct parent of GO:0071816; redundant general term.
    supported_by:
    - reference_id: PMID:23041287
      supporting_text: an ATPase targeting newly synthesized TA proteins
- term:
    id: GO:0071816
    label: tail-anchored membrane protein insertion into ER membrane
  evidence_type: IMP
  original_reference_id: PMID:31461301
  qualifier: involved_in
  review:
    summary: Mutant-phenotype evidence that the disease-associated Val163Ala ASNA1 mutant, while still able to capture a TA substrate, is inefficient in facilitating TA insertion into the ER membrane; asna1-null zebrafish show cardiac failure. Establishes GET3's role in the TA-insertion pathway.
    action: ACCEPT
    reason: Core biological process with mutant-phenotype (IMP) support directly linking ASNA1 function to TA insertion.
    supported_by:
    - reference_id: PMID:31461301
      supporting_text: inefficient in facilitating TA protein insertion into the ER membrane
- term:
    id: GO:0140597
    label: protein carrier activity
  evidence_type: IDA
  original_reference_id: PMID:23610396
  qualifier: enables
  review:
    summary: GET3/TRC40 is the TMD chaperone/carrier that shields the hydrophobic TA transmembrane domain and delivers it to the ER, harnessing ATP to drive TA membrane localization. This protein-carrier (chaperone) activity is a core molecular function complementary to its ATPase activity.
    action: ACCEPT
    reason: Core molecular function; GET3 carries the TA-protein cargo as a soluble TMD chaperone, demonstrated by IDA.
    supported_by:
    - reference_id: PMID:23610396
      supporting_text: Get3 harnesses the energy from ATP to drive
    - reference_id: PMID:37963916
      supporting_text: the Get3 chaperone captures the TA protein substrate and delivers it to the Get1/Get2 membrane protein complex (GET insertase)
- term:
    id: GO:0043529
    label: GET complex
  evidence_type: IDA
  original_reference_id: PMID:32910895
  qualifier: part_of
  review:
    summary: Direct structural evidence (cryo-EM) placing GET3/TRC40 within the GET insertase complex.
    action: ACCEPT
    reason: Core cellular component; structurally demonstrated.
    supported_by:
    - reference_id: PMID:32910895
      supporting_text: captures the TA from a cytoplasmic chaperone (Get3 or TRC40, respectively)
- term:
    id: GO:0071816
    label: tail-anchored membrane protein insertion into ER membrane
  evidence_type: IMP
  original_reference_id: PMID:23041287
  qualifier: involved_in
  review:
    summary: Mutant-phenotype evidence supporting GET3/TRC40's role as the ATPase targeting TA proteins for insertion, delivered to the CAML/WRB receptor complex.
    action: ACCEPT
    reason: Core biological process with IMP support.
    supported_by:
    - reference_id: PMID:23041287
      supporting_text: an ATPase targeting newly synthesized TA proteins
- term:
    id: GO:0043529
    label: GET complex
  evidence_type: IPI
  original_reference_id: PMID:23041287
  qualifier: part_of
  review:
    summary: IPI identification of GET3/TRC40 in the receptor (GET) complex with WRB and CAML.
    action: ACCEPT
    reason: Core cellular component; demonstrated by complex identification.
    supported_by:
    - reference_id: PMID:23041287
      supporting_text: CAML and WRB as components of the TRC40 receptor complex
- term:
    id: GO:0071816
    label: tail-anchored membrane protein insertion into ER membrane
  evidence_type: IDA
  original_reference_id: PMID:25535373
  qualifier: involved_in
  review:
    summary: Direct evidence that the minimal Bag6 complex facilitates TA substrate transfer from SGTA to TRC40/GET3, the loading step that commits TA proteins to the GET targeting pathway.
    action: ACCEPT
    reason: Core biological process; IDA demonstrating the TA-loading step onto GET3.
    supported_by:
    - reference_id: PMID:25535373
      supporting_text: substrate transfer from small glutamine-rich tetratricopeptide repeat-containing
- term:
    id: GO:0070062
    label: extracellular exosome
  evidence_type: HDA
  original_reference_id: PMID:19056867
  qualifier: located_in
  review:
    summary: High-throughput urinary exosome proteomics catalog hit. Not indicative of a core localization or function for GET3.
    action: KEEP_AS_NON_CORE
    reason: Proteomic catalog localization; not part of GET3's core cytosolic targeting function.
    supported_by:
    - reference_id: PMID:19056867
      supporting_text: Large-scale proteomics and phosphoproteomics of urinary exosomes
- term:
    id: GO:0005730
    label: nucleolus
  evidence_type: TAS
  original_reference_id: PMID:9736449
  qualifier: located_in
  review:
    summary: Legacy nucleolar localization reported in the early arsenite-ATPase characterization (cytoplasmic, perinuclear, and nucleolar distribution). Secondary to the core cytosolic targeting function.
    action: KEEP_AS_NON_CORE
    reason: Legacy localization from early arsA-homolog work; not part of the core GET-pathway function.
    supported_by:
    - reference_id: PMID:9736449
      supporting_text: Dual cytoplasmic and nuclear distribution of the novel arsenite-stimulated human ATPase
- term:
    id: GO:0005737
    label: cytoplasm
  evidence_type: TAS
  original_reference_id: PMID:9736449
  qualifier: located_in
  review:
    summary: Cytoplasmic localization from the early arsenite-ATPase study, consistent with GET3's primary cytosolic site of action.
    action: ACCEPT
    reason: Correct primary compartment; corroborated by experimental EXP evidence.
    supported_by:
    - reference_id: PMID:9736449
      supporting_text: Dual cytoplasmic and nuclear distribution
references:
- id: GO_REF:0000033
  title: Annotation inferences using phylogenetic trees
  findings: []
- id: GO_REF:0000044
  title: Gene Ontology annotation based on UniProtKB/Swiss-Prot Subcellular Location vocabulary mapping
  findings: []
- id: GO_REF:0000052
  title: Gene Ontology annotation based on curation of immunofluorescence data
  findings: []
- id: GO_REF:0000104
  title: Electronic Gene Ontology annotations created by transferring manual GO annotations between related proteins based on shared sequence features
  findings: []
- id: GO_REF:0000120
  title: Combined Automated Annotation using Multiple IEA Methods
  findings: []
- id: PMID:17382883
  title: Identification of a targeting factor for posttranslational membrane protein insertion into the ER.
  findings:
  - statement: Identified the cytosolic TMD recognition complex (TRC); the 40 kDa ATPase subunit TRC40 is Asna-1, which targets TA proteins for ER insertion with release dependent on ATP hydrolysis.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Founding study identifying TRC40/Asna1 (GET3) as the cytosolic TA-targeting ATPase.
- id: PMID:19056867
  title: Large-scale proteomics and phosphoproteomics of urinary exosomes.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: High-throughput exosome proteomics; source of the extracellular exosome catalog annotation.
- id: PMID:21444755
  title: WRB is the receptor for TRC40/Asna1-mediated insertion of tail-anchored proteins into the ER membrane.
  findings:
  - statement: TRC40/Asna1 (Get3 in yeast) is a conserved cytosolic ATPase that recognizes the TMD of TA proteins and delivers them to the ER membrane receptor WRB.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Establishes GET3/TRC40 as the cytosolic ATPase delivering TA proteins to the ER WRB receptor.
- id: PMID:21516116
  title: Next-generation sequencing to generate interactome datasets.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: High-throughput interactome; bare protein binding source.
- id: PMID:21911467
  title: Insight into bacterial virulence mechanisms against host immune response via the Yersinia pestis-human protein-protein interaction network.
  findings: []
  reference_review:
    relevance: NONE
    correctness: VERIFIED
    review_notes: Host-pathogen PPI screen; the GET3 partner is a bacterial protein (yscD), incidental and not relevant to GET3 function.
- id: PMID:23041287
  title: Molecular machinery for insertion of tail-anchored membrane proteins into the endoplasmic reticulum membrane in mammalian cells.
  findings:
  - statement: CAML and WRB are components of the TRC40 receptor complex; TRC40 is an ATPase targeting newly synthesized TA proteins to the ER for insertion.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Defines the mammalian TRC40 receptor complex and GET3's targeting role.
- id: PMID:23610396
  title: Precise timing of ATPase activation drives targeting of tail-anchored proteins.
  findings:
  - statement: Get3 coordinates delivery of TA proteins to the ER; the Get4/5 loading complex locks Get3 in the ATP-bound state while the TA substrate activates Get3's ATPase ~100-fold, and Get3 harnesses ATP energy to drive TA membrane localization.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Mechanistic study of the Get3 ATPase/carrier cycle; supports protein carrier (chaperone) activity.
- id: PMID:25416956
  title: A proteome-scale map of the human interactome network.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: High-throughput interactome; bare protein binding source.
- id: PMID:25535373
  title: Bag6 complex contains a minimal tail-anchor-targeting module and a mock BAG domain.
  findings:
  - statement: The minimal Bag6 complex facilitates TA substrate transfer from SGTA to TRC40, the loading step of the GET pathway.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Establishes the SGTA-to-TRC40/GET3 TA handoff.
- id: PMID:28514442
  title: Architecture of the human interactome defines protein communities and disease networks.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: High-throughput interactome; bare protein binding, includes CAMLG.
- id: PMID:31461301
  title: Biallelic Variants in ASNA1, Encoding a Cytosolic Targeting Factor of Tail-Anchored Proteins, Cause Rapidly Progressive Pediatric Cardiomyopathy.
  findings:
  - statement: ASNA1 (TRC40/GET3) is a ubiquitously expressed cytosolic chaperone mediating TA-protein insertion into the ER; the disease Val163Ala mutant captures TA but is inefficient in facilitating insertion, and asna1-null zebrafish develop cardiac failure.
    reference_section_type: RESULTS
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Disease study; provides IMP evidence linking ASNA1/GET3 to TA insertion and human cardiomyopathy.
- id: PMID:32296183
  title: A reference map of the human binary protein interactome.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: HuRI Y2H interactome; bare protein binding source.
- id: PMID:32910895
  title: Structural Basis of Tail-Anchored Membrane Protein Biogenesis by the GET Insertase Complex.
  findings:
  - statement: Cryo-EM of the GET insertase complex; the WRB/CAML (Get1/Get2) insertase captures the TA from the cytoplasmic chaperone GET3/TRC40 that GET3 binds as a homodimer.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Definitive structure showing GET3 docking on the WRB/CAML insertase.
- id: PMID:33961781
  title: Dual proteome-scale networks reveal cell-specific remodeling of the human interactome.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: High-throughput interactome; bare protein binding, includes CAMLG.
- id: PMID:40205054
  title: Multimodal cell maps as a foundation for structural and functional genomics.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: High-throughput cell map; bare protein binding, includes CAMLG.
- id: PMID:8884272
  title: Isolation of the ATP-binding human homolog of the arsA component of the bacterial arsenite transporter.
  findings:
  - statement: Isolated the human homolog of bacterial arsA (hARSA-I/ASNA1) and showed it is an ATPase analogous to bacterial ArsA.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: MEDIUM
    correctness: VERIFIED
    review_notes: Original cloning of ASNA1 as an arsA homolog; basis of the ATPase MF, legacy arsenite framing.
- id: PMID:9712828
  title: Biochemical characterization of the human arsenite-stimulated ATPase (hASNA-I).
  findings:
  - statement: Recombinant hASNA-I exhibits basal ATPase activity (modestly arsenite-stimulated; antimonite does not stimulate), with measured ATP kinetics.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: MEDIUM
    correctness: VERIFIED
    review_notes: Biochemical ATPase characterization; legacy arsenite-stimulation is secondary to the TA-targeting role.
- id: PMID:9736449
  title: Dual cytoplasmic and nuclear distribution of the novel arsenite-stimulated human ATPase (hASNA-I).
  findings:
  - statement: Early study reporting a cytoplasmic, perinuclear, and nucleolar distribution of the arsenite-stimulated ATPase.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: Source of legacy nucleolar/cytoplasmic localization annotations.
- id: PMID:34264263
  title: Capture and delivery of tail-anchored proteins to the endoplasmic reticulum.
  findings:
  - statement: Comprehensive review of the GET/TRC pathway; GET3/TRC40 is the central cytosolic homodimeric ATPase that captures TA substrates, shields the hydrophobic TMD, and delivers them to the ER WRB/CAML receptor-insertase for insertion.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: PubMed-verified (J Cell Biol 2021, 220:8). Authoritative pathway review of GET3/TRC40 as the cytosolic targeting ATPase/TMD chaperone.
- id: PMID:36640319
  title: The Get1/2 insertase forms a channel to mediate the insertion of tail-anchored proteins into the ER.
  findings:
  - statement: Get3 (TRC40) delivers TA proteins to the Get1/2 channel; Get3 binding seals the dynamically opening Get1/2 channel, and channel activity is required to release TA proteins from Get3 for insertion, defining the GET3-to-insertase handoff step.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: MEDIUM
    correctness: VERIFIED
    review_notes: PubMed-verified (Cell Rep 2022, 42:111921). Mechanistic context for GET3/TRC40 handing off TA cargo to the Get1/2 (WRB/CAML) insertase channel; Get3 gates the channel.
- id: PMID:37963916
  title: The GET insertase exhibits conformational plasticity and induces membrane thinning.
  findings:
  - statement: Structures and simulations of human and C. thermophilum Get1/Get2/Get3 show that the gating interaction between Get2 helix alpha3' and Get3 drives conformational changes in both Get3 and the Get1/Get2 heterotetramer, promoting Get3 opening, nucleotide release, and TA substrate transfer to the membrane.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: PubMed-verified (Nat Commun 2023, 14:7355). Structural mechanism of the Get3/Get1/Get2 cycle including GET3/TRC40 conformational opening on receptor engagement; includes human GET3.
- id: file:human/GET3/GET3-uniprot.txt
  title: UniProt entry O43681 (GET3_HUMAN), ATPase ASNA1/TRC40
  findings:
  - statement: ATPase required for post-translational delivery of TA proteins to the ER; recognizes the TA TMD in the cytosol, targets to the GET1/WRB-CAMLG/GET2 receptor where ATP hydrolysis drives release/insertion; component of the GET complex; cytoplasmic/ER/nucleolar.
    reference_section_type: OTHER
core_functions:
- description: Cytosolic ATPase and TMD chaperone of the GET/TRC pathway that recognizes and binds the C-terminal transmembrane domain of newly synthesized tail-anchored proteins and, in an ATP-driven cycle, carries and delivers them to the ER membrane receptor for insertion.
  molecular_function:
    id: GO:0016887
    label: ATP hydrolysis activity
  locations:
  - id: GO:0005737
    label: cytoplasm
  supported_by:
  - reference_id: PMID:17382883
    supporting_text: cytosolic TMD recognition complex (TRC) that targets TA proteins for insertion into the ER membrane
  - reference_id: file:human/GET3/GET3-uniprot.txt
    supporting_text: ATP hydrolysis is required for insertion.
  directly_involved_in:
  - id: GO:0071816
    label: tail-anchored membrane protein insertion into ER membrane
- description: Soluble protein carrier (TMD chaperone) that shields the hydrophobic tail anchor of TA substrates and, as part of the GET complex docking on the WRB/CAML insertase, delivers them for ER membrane insertion.
  molecular_function:
    id: GO:0140597
    label: protein carrier activity
  in_complex:
    id: GO:0043529
    label: GET complex
  supported_by:
  - reference_id: PMID:23610396
    supporting_text: Get3 harnesses the energy from ATP to drive
  - reference_id: file:human/GET3/GET3-uniprot.txt
    supporting_text: Recognizes and selectively binds the transmembrane domain of TA
  directly_involved_in:
  - id: GO:0071816
    label: tail-anchored membrane protein insertion into ER membrane
proposed_new_terms: []
suggested_questions:
- question: To what extent is the legacy arsenite/antimonite-stimulated ATPase activity physiologically relevant in humans, versus being a vestige of the ancestral ArsA fold now repurposed for TA targeting?
- question: Which specific cardiac tail-anchored proteins are mislocalized when ASNA1 is impaired, and do they account for the dilated cardiomyopathy phenotype?
suggested_experiments:
- description: Reconstitute the full handoff cascade (SGTA -> BAG6/UBL4A/GET4 -> GET3 -> WRB/CAML) with purified components to quantify how GET4 priming and substrate-induced ATPase activation set the timing of TA capture and release.
- description: Define the endogenous GET3/ASNA1 TA-substrate repertoire in cardiomyocytes by proximity labeling and compare wild-type versus the Val163Ala disease variant to identify the TA clients whose mislocalization drives cardiomyopathy.

GET3

Gene Description

Existing Annotations Review

Core Functions

Cytosolic ATPase and TMD chaperone of the GET/TRC pathway that recognizes and binds the C-terminal transmembrane domain of newly synthesized tail-anchored proteins and, in an ATP-driven cycle, carries and delivers them to the ER membrane receptor for insertion.

Soluble protein carrier (TMD chaperone) that shields the hydrophobic tail anchor of TA substrates and, as part of the GET complex docking on the WRB/CAML insertase, delivers them for ER membrane insertion.

References

Suggested Questions for Experts

Suggested Experiments

Deep Research

Falcon

Research report: Human GET3 / ASNA1 / TRC40 (UniProt O43681)

0) Target verification (gene/protein identity)

1) Key concepts and definitions (current understanding)

Tail-anchored (TA) proteins

GET/TRC pathway (ER targeting route for TA proteins)

ASNA1/TRC40/GET3 (central ATPase/chaperone)

2) Molecular function and biochemical mechanism

2.1 Enzymatic activity

2.2 Substrate specificity (what does ASNA1/TRC40 act on?)

2.3 Stepwise pathway model (from synthesis to ER insertion)

Mechanistic detail from 2023 structural work

3) Cellular localization (where does ASNA1/TRC40 function?)

4) Recent developments (prioritizing 2023–2024)

4.1 2023: GET insertase structural plasticity and membrane remodeling

4.2 2023: A channel model for Get1/2-mediated insertion

4.3 2023: Updated reviews of TA targeting and surveillance

4.4 2024: Redox-regulated functional switching (TRC40 as a stress chaperone)

5) Current applications and real-world implementations

5.1 Functional genomics and proteomics “client spectrum” mapping at the ER

5.2 Stress biology / proteostasis (emerging)

5.3 Quality control implications for TA biogenesis

6) Expert opinions and analysis (authoritative interpretation)

7) Relevant statistics and quantitative data points (from recent studies)

8) Notes on “arsenite-stimulated ATPase” nomenclature

9) Summary for functional annotation (human UniProt O43681)

Evidence synopsis table

Artifacts

Citations

📚 Additional Documentation

Notes

GET3 (ASNA1 / TRC40) — review notes

Core function: cytosolic ATPase that TARGETS tail-anchored (TA) proteins to the ER

ATPase / ATP binding (MF)

Protein carrier / chaperone activity (MF)

GET complex / receptor (CC + targeting)

Pre-targeting / bridging complex interactions (IPI partners)

Subcellular location

Legacy / secondary role: arsenite-stimulated ATPase (arsA homolog)

Disease

Annotation decisions summary

Falcon deep-research findings (incorporated 2026-06)

Pn Notes

GET3 PN Consistency Notes

Source Files Checked

Deep Research Files

AIGR Review Snapshot

PN Consistency Summary

Full Consistency Review

PN Dossier Context

PN row 1: ER proteostasis | Protein transport | GET pathway component

Projected GO annotations (2)

Note

📄 View Raw YAML