id: Q9H8Y5 gene_symbol: ANKZF1 product_type: PROTEIN status: COMPLETE taxon: id: NCBITaxon:9606 label: Homo sapiens description: >- ANKZF1 is a cytosolic tRNA endonuclease in the ribosome-associated quality-control pathway. It acts after stalled-ribosome splitting on 60S ribosome-nascent-chain complexes, cleaving the terminal 3'-CCA region of P-site peptidyl-tRNA to release incomplete nascent polypeptides for degradation and to generate tRNA repair intermediates that can be recycled. ANKZF1 also binds VCP/p97 and has a stress-responsive mitochondrial context, translocating to mitochondria under oxidative stress and supporting mitochondrial integrity. 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, accompanied by conservative changes to GO terms applied by UniProt findings: [] - id: PMID:19946888 title: Defining the membrane proteome of NK cells. findings: [] - id: PMID:22190034 title: Global landscape of HIV-human protein complexes. findings: [] - id: PMID:28302725 title: Ankyrin repeat and zinc-finger domain-containing 1 mutations are associated with infantile-onset inflammatory bowel disease. findings: [] - id: PMID:29632312 title: Vms1 and ANKZF1 peptidyl-tRNA hydrolases release nascent chains from stalled ribosomes. findings: [] - id: PMID:30244831 title: Release of Ubiquitinated and Non-ubiquitinated Nascent Chains from Stalled Mammalian Ribosomal Complexes by ANKZF1 and Ptrh1. findings: [] - id: PMID:31011209 title: Mechanism for recycling tRNAs on stalled ribosomes. findings: [] - id: PMID:32075755 title: ELAC1 Repairs tRNAs Cleaved during Ribosome-Associated Quality Control. findings: [] - id: PMID:21896481 title: The general definition of the p97/valosin-containing protein (VCP)-interacting motif (VIM) delineates a new family of p97 cofactors. full_text_unavailable: true findings: - statement: >- Defines the VCP-interacting motif (VIM, minimal consensus RX5AAX2R) and identifies ZNF744/ANKZF1 as a member of the VIM-containing p97/VCP cofactor family; the VIM is necessary and sufficient for binding the p97 N-terminal domain. - id: PMID:38412259 title: Ribosomal quality control factors inhibit repeat-associated non-AUG translation from GC-rich repeats. full_text_unavailable: true findings: - statement: >- Depletion of the RQC factors NEMF, LTN1 and ANKZF1 increases accumulation of repeat-associated non-AUG (RAN) translation products from G4C2 (C9ORF72) and CGG (FMR1) repeats, while their overexpression reduces RAN products in reporters and in C9ALS/FTD patient iPSC-derived neurons. - id: PMID:38388640 title: Stalled translation by mitochondrial stress upregulates a CNOT4-ZNF598 ribosomal quality control pathway important for tissue homeostasis. full_text_unavailable: true findings: - statement: >- Mitochondrial stress upregulates a CNOT4-ZNF598 ribosome-associated quality-control pathway whose downstream steps include ANKZF1-mediated release of stalled nascent chains, linking RQC to mitochondrial and tissue homeostasis. - id: PMID:37158785 title: Human ESC-derived vascular cells promote vascular regeneration in a HIF-1alpha dependent manner. full_text_unavailable: true findings: - statement: >- ANKZF1 is reported as a HIF-1alpha effector; transcriptional inactivation of ANKZF1 in human mesenchymal stem cells impairs pro-angiogenic processes under hypoxia. - id: PMID:31257922 title: High ANKZF1 expression is associated with poor overall survival and recurrence-free survival in colon cancer. full_text_unavailable: true findings: - statement: >- A TCGA bioinformatic analysis reports that high ANKZF1 expression is associated with poorer overall survival and recurrence-free survival in colon cancer; this is a correlative expression-outcome association. - id: Reactome:R-HSA-9948299 title: Ribosome-associated quality control findings: [] - id: Reactome:R-HSA-9948427 title: ANKZF1 cleaves the peptidyl-tRNA in VCP hexamer:LTN1:NEMF:60S ribosome subunit:peptidyl-tRNA with K48polyUb-nascent peptide findings: [] - id: file:human/ANKZF1/ANKZF1-uniprot.txt title: UniProt record for human ANKZF1 findings: [] - id: file:human/ANKZF1/ANKZF1-notes.md title: ANKZF1 curation notes findings: [] - id: file:projects/PROTEOSTASIS/reports/pn_projection/pn_projected_gene_go_summary.tsv title: Proteostasis Network projected GO summary findings: [] - id: file:projects/PROTEOSTASIS/reports/pn_projection/pn_projected_candidate_additions.tsv title: Proteostasis Network projected candidate additions findings: [] - id: file:projects/PROTEOSTASIS/mappings/mitochondrial_proteostasis.yaml title: Proteostasis Network mitochondrial proteostasis mappings findings: [] - id: file:projects/PROTEOSTASIS/reports/pn_mapping_audit/current_mapping_scrutiny.tsv title: Proteostasis Network mapping audit findings: [] existing_annotations: - term: id: GO:0036503 label: ERAD pathway evidence_type: IBA original_reference_id: GO_REF:0000033 qualifier: involved_in review: summary: >- Remove. The phylogenetic transfer appears to over-project a Vms1/Cdc48 quality-control relationship into ERAD. Human ANKZF1 evidence supports cytosolic ribosome-associated quality control and oxidative-stress mitochondrial context, not direct endoplasmic-reticulum-associated degradation. action: REMOVE reason: >- The experimentally supported conserved function is cleavage of peptidyl-tRNA on 60S RQC complexes, with downstream nascent-chain degradation. That is distinct from the ERAD pathway. additional_reference_ids: - PMID:30244831 - PMID:31011209 - PMID:32075755 supported_by: - reference_id: PMID:30244831 supporting_text: ANKZF1 does not function as a peptidyl-tRNA hydrolase. - reference_id: PMID:31011209 supporting_text: ANKZF1 and Vms1p sever - term: id: GO:0005737 label: cytoplasm evidence_type: IEA original_reference_id: GO_REF:0000044 qualifier: located_in review: summary: >- Correct broad localization. UniProt and the oxidative-stress study place ANKZF1 in the cytoplasm, with stress-dependent mitochondrial translocation. action: ACCEPT reason: >- Cytoplasm is a valid broad cellular component for ANKZF1, although the core RQC activity is more specifically cytosolic. additional_reference_ids: - PMID:28302725 - file:human/ANKZF1/ANKZF1-uniprot.txt supported_by: - reference_id: PMID:28302725 supporting_text: ANKZF1 is located diffusely in the cytoplasm - term: id: GO:0005515 label: protein binding evidence_type: IPI original_reference_id: PMID:22190034 qualifier: enables review: summary: >- Over-annotated. The source is a large-scale HIV-host interaction study and the generic protein binding term does not describe ANKZF1's biochemical role. action: MARK_AS_OVER_ANNOTATED reason: >- Generic protein binding is uninformative for ANKZF1. A biologically relevant interaction is VCP/p97 binding through the VIM motif (consensus RX5AAX2R, defined in Stapf et al. 2011, which identified ANKZF1/ZNF744 as a VIM-containing p97 cofactor), but this HIV AP-MS annotation should not be treated as core function. additional_reference_ids: - PMID:21896481 - file:human/ANKZF1/ANKZF1-uniprot.txt supported_by: - reference_id: PMID:22190034 supporting_text: physical interactions of all 18 HIV-1 proteins and polyproteins with host - term: id: GO:0072344 label: rescue of stalled cytosolic ribosome evidence_type: TAS original_reference_id: Reactome:R-HSA-9948299 qualifier: involved_in review: summary: >- Correct. Reactome places ANKZF1 in the RQC pathway, where it cleaves the tRNA portion of peptidyl-tRNA on stalled-ribosome 60S complexes. action: ACCEPT reason: >- Rescue of stalled cytosolic ribosomes is a core biological process for ANKZF1 because its tRNA cleavage releases nascent chains from 60S RQC complexes. supported_by: - reference_id: Reactome:R-HSA-9948299 supporting_text: ANKZF1, which interacts with VCP, cleaves the C-terminal - term: id: GO:0004521 label: RNA endonuclease activity evidence_type: TAS original_reference_id: Reactome:R-HSA-9948427 qualifier: enables review: summary: >- The activity is correct but the more specific term tRNA-specific ribonuclease activity better captures ANKZF1's substrate and reaction. action: MODIFY reason: >- Reactome describes cleavage of the tRNA portion of peptidyl-tRNA. Use the more specific tRNA-specific ribonuclease activity term rather than generic RNA endonuclease activity. proposed_replacement_terms: - id: GO:0004549 label: tRNA-specific ribonuclease activity supported_by: - reference_id: Reactome:R-HSA-9948427 supporting_text: ANKZF1 cleaves the C-terminal 3 nucleotides, CCA, of the tRNA - term: id: GO:0005829 label: cytosol evidence_type: TAS original_reference_id: Reactome:R-HSA-9948427 qualifier: located_in review: summary: >- Correct. The RQC substrate and Reactome event are cytosolic ribosome-nascent-chain complexes. action: ACCEPT reason: >- Cytosol is the best core cellular component for the RQC tRNA endonuclease activity. additional_reference_ids: - PMID:30244831 - PMID:32075755 supported_by: - reference_id: PMID:32075755 supporting_text: Ribosome-associated quality control (RQC) disassembles aberrantly stalled - term: id: GO:0004521 label: RNA endonuclease activity evidence_type: IDA original_reference_id: PMID:30244831 qualifier: enables review: summary: >- The experimental activity is correct but should be represented by the more specific tRNA-specific ribonuclease activity term. action: MODIFY reason: >- Kuroha et al. showed that ANKZF1 induces specific cleavage in the acceptor arm of 60S-bound P-site tRNA, so a tRNA-specific ribonuclease term is more informative than generic RNA endonuclease activity. proposed_replacement_terms: - id: GO:0004549 label: tRNA-specific ribonuclease activity supported_by: - reference_id: PMID:30244831 supporting_text: specific cleavage in the acceptor arm of 60S-bound P site tRNA - term: id: GO:0004521 label: RNA endonuclease activity evidence_type: IDA original_reference_id: PMID:31011209 qualifier: enables review: summary: >- The experimental activity is correct but should be represented by the more specific tRNA-specific ribonuclease activity term. action: MODIFY reason: >- The paper identifies precise cleavage of the terminal 3'CCA nucleotides of polypeptidyl-tRNAs on RQC complexes, matching tRNA-specific ribonuclease activity more closely than generic RNA endonuclease activity. proposed_replacement_terms: - id: GO:0004549 label: tRNA-specific ribonuclease activity supported_by: - reference_id: PMID:31011209 supporting_text: polypeptidyl-tRNAs on RQC complexes by precisely cleaving off the terminal 3'CCA - term: id: GO:0006515 label: protein quality control for misfolded or incompletely synthesized proteins evidence_type: IDA original_reference_id: PMID:29632312 qualifier: involved_in review: summary: >- Correct. ANKZF1 acts in ribosome-associated quality control to release nascent chains from stalled ribosomes for degradation. action: ACCEPT reason: >- This term captures the protein-quality-control outcome of ANKZF1's RQC activity. supported_by: - reference_id: PMID:29632312 supporting_text: Failure to degrade the NCs leads to protein aggregation and proteotoxic stress - term: id: GO:0006515 label: protein quality control for misfolded or incompletely synthesized proteins evidence_type: IDA original_reference_id: PMID:30244831 qualifier: involved_in review: summary: >- Correct. The mammalian RQC reconstitution links ANKZF1-mediated tRNA cleavage to release of nascent chains that can be degraded by the proteasome. action: ACCEPT reason: >- ANKZF1 directly supports quality control of aberrant translation products in RQC. supported_by: - reference_id: PMID:30244831 supporting_text: In conclusion, ANKZF1 induces specific tRNA cleavage in ubiquitinated 60S RNCs - term: id: GO:0006515 label: protein quality control for misfolded or incompletely synthesized proteins evidence_type: IDA original_reference_id: PMID:31011209 qualifier: involved_in review: summary: >- Correct. Cleavage of polypeptidyl-tRNAs on RQC complexes releases ubiquitinated nascent proteins for proteasomal degradation. action: ACCEPT reason: >- The activity is a core protein quality-control step for incompletely synthesized polypeptides on stalled ribosomes. supported_by: - reference_id: PMID:31011209 supporting_text: releases ubiquitinated nascent proteins from 60S ribosomal subunits for proteasomal - term: id: GO:0006515 label: protein quality control for misfolded or incompletely synthesized proteins evidence_type: IDA original_reference_id: PMID:32075755 qualifier: involved_in review: summary: >- Correct. ELAC1 repair work confirms ANKZF1-dependent tRNA cleavage during ribosome stalling in mammalian cells. action: ACCEPT reason: >- The paper supports ANKZF1 as a cellular RQC factor whose cleavage products are repaired and recycled after stalled-ribosome events. supported_by: - reference_id: PMID:32075755 supporting_text: Deleting ELAC1 leads to the ANKZF1-dependent accumulation of unrepaired tRNA intermediates - term: id: GO:0072344 label: rescue of stalled cytosolic ribosome evidence_type: IDA original_reference_id: PMID:29632312 qualifier: involved_in review: summary: >- Correct. Purified human ANKZF1 can act on stalled-ribosome peptidyl-tRNA substrates in the conserved RQC release step. action: ACCEPT reason: >- The annotation captures the RQC/ribosome-rescue context of the ANKZF1 activity. supported_by: - reference_id: PMID:29632312 supporting_text: Purified Ankzf1 - term: id: GO:0072344 label: rescue of stalled cytosolic ribosome evidence_type: IDA original_reference_id: PMID:30244831 qualifier: involved_in review: summary: >- Correct. In vitro mammalian RQC reconstitution shows ANKZF1 acting on 60S-bound P-site tRNA after stalled-ribosome processing. action: ACCEPT reason: >- ANKZF1-mediated cleavage of 60S RQC complexes is a direct stalled-ribosome rescue step. supported_by: - reference_id: PMID:30244831 supporting_text: Here, we reconstituted the mammalian RQC pathway in vitro - term: id: GO:0072344 label: rescue of stalled cytosolic ribosome evidence_type: IDA original_reference_id: PMID:31011209 qualifier: involved_in review: summary: >- Correct. ANKZF1 liberates peptidyl-tRNAs from stalled ribosomes by cleaving the terminal 3'CCA region. action: ACCEPT reason: >- This is a direct mechanistic description of the stalled-ribosome rescue role. supported_by: - reference_id: PMID:31011209 supporting_text: ANKZF1 liberates peptidyl-tRNAs - term: id: GO:0072344 label: rescue of stalled cytosolic ribosome evidence_type: IDA original_reference_id: PMID:32075755 qualifier: involved_in review: summary: >- Correct. The ELAC1 study describes ANKZF1 cleavage of P-site tRNA as a key RQC step after ribosome stalling. action: ACCEPT reason: >- The annotation is directly supported by stalled-ribosome RQC evidence. supported_by: - reference_id: PMID:32075755 supporting_text: RQC is the cleavage of P-site tRNA by the endonuclease ANKZF1 - term: id: GO:0140101 label: catalytic activity, acting on a tRNA evidence_type: IDA original_reference_id: PMID:30244831 qualifier: enables review: summary: >- Correct substrate class but too broad. ANKZF1's activity is specifically tRNA ribonuclease cleavage in the 60S RQC substrate context. action: MODIFY reason: >- Replace the generic tRNA catalytic-activity term with tRNA-specific ribonuclease activity. proposed_replacement_terms: - id: GO:0004549 label: tRNA-specific ribonuclease activity supported_by: - reference_id: PMID:30244831 supporting_text: ANKZF1 induces specific tRNA cleavage in ubiquitinated 60S RNCs - term: id: GO:0140101 label: catalytic activity, acting on a tRNA evidence_type: IDA original_reference_id: PMID:31011209 qualifier: enables review: summary: >- Correct substrate class but too broad. The paper supports precise tRNA cleavage rather than an unspecified tRNA catalytic activity. action: MODIFY reason: >- Replace the generic tRNA catalytic-activity term with tRNA-specific ribonuclease activity. proposed_replacement_terms: - id: GO:0004549 label: tRNA-specific ribonuclease activity supported_by: - reference_id: PMID:31011209 supporting_text: terminal 3'CCA nucleotides universal to all tRNAs - term: id: GO:0140101 label: catalytic activity, acting on a tRNA evidence_type: IDA original_reference_id: PMID:32075755 qualifier: enables review: summary: >- Correct substrate class but too broad. ANKZF1 is a tRNA endonuclease in RQC, not merely a generic enzyme acting on tRNA. action: MODIFY reason: >- Replace the generic tRNA catalytic-activity term with tRNA-specific ribonuclease activity. proposed_replacement_terms: - id: GO:0004549 label: tRNA-specific ribonuclease activity supported_by: - reference_id: PMID:32075755 supporting_text: tRNAs cleaved by ANKZF1 for CCA re-addition - term: id: GO:0005737 label: cytoplasm evidence_type: IDA original_reference_id: PMID:28302725 qualifier: located_in review: summary: >- Correct. The disease/stress paper reports diffuse cytoplasmic localization with translocation to mitochondria under cellular stress. action: ACCEPT reason: >- Cytoplasm is experimentally supported, though the main RQC activity is cytosolic. supported_by: - reference_id: PMID:28302725 supporting_text: ANKZF1 is located diffusely in the cytoplasm - term: id: GO:0070301 label: cellular response to hydrogen peroxide evidence_type: IMP original_reference_id: PMID:28302725 qualifier: involved_in review: summary: >- Supported but non-core. ANKZF1 translocates to mitochondria during oxidative stress and loss of ANKZF1 affects mitochondrial integrity and respiration under stress. action: KEEP_AS_NON_CORE reason: >- Hydrogen-peroxide response is an experimentally observed stress phenotype and localization context, but the best-supported core molecular function is RQC-associated tRNA endonuclease activity. additional_reference_ids: - file:human/ANKZF1/ANKZF1-uniprot.txt supported_by: - reference_id: PMID:28302725 supporting_text: translocates to the mitochondria upon cellular stress - term: id: GO:0016020 label: membrane evidence_type: HDA original_reference_id: PMID:19946888 qualifier: located_in review: summary: >- Over-annotated. The source is a high-throughput NK-cell membrane proteome and does not establish ANKZF1 as a membrane protein. action: MARK_AS_OVER_ANNOTATED reason: >- UniProt and direct ANKZF1 studies support cytoplasmic/cytosolic localization with stress-dependent mitochondrial translocation, not stable membrane residence. additional_reference_ids: - PMID:28302725 - file:human/ANKZF1/ANKZF1-uniprot.txt supported_by: - reference_id: PMID:19946888 supporting_text: transiently associated with membranes core_functions: - description: >- ANKZF1 cleaves the tRNA moiety of P-site peptidyl-tRNAs on 60S ribosome-nascent-chain complexes in ribosome-associated quality control, allowing incomplete nascent polypeptides to be released for proteasomal degradation and ANKZF1-cleaved tRNAs to enter repair and recycling pathways. molecular_function: id: GO:0004549 label: tRNA-specific ribonuclease activity directly_involved_in: - id: GO:0072344 label: rescue of stalled cytosolic ribosome - id: GO:0006515 label: protein quality control for misfolded or incompletely synthesized proteins locations: - id: GO:0005829 label: cytosol supported_by: - reference_id: PMID:30244831 supporting_text: specific cleavage in the acceptor arm of 60S-bound P site tRNA - reference_id: PMID:31011209 supporting_text: polypeptidyl-tRNAs on RQC complexes by precisely cleaving off the terminal 3'CCA - reference_id: PMID:32075755 supporting_text: tRNAs cleaved by ANKZF1 for CCA re-addition proposed_new_terms: [] suggested_questions: - question: >- Does human ANKZF1 directly drive catabolism of mitochondrial proteins, or is the mitochondrial connection limited to stress-dependent localization and mitochondrial integrity phenotypes downstream of its RQC/VCP biology? experts: - Siu Sylvia Lee - Shin-ichi Kuroha - Sichen Shao - question: >- In mammalian cells, does ANKZF1 act on mitochondria-targeted stalled nascent chains through the same cytosolic RQC substrate state, or through a distinct organelle-proximal mechanism? experts: - Siu Sylvia Lee - Sichen Shao - question: >- Is the VCP/p97 interaction via the ANKZF1 VIM motif (consensus RX5AAX2R) required for ANKZF1's RQC nascent-chain release activity in cells, and should ANKZF1 carry a specific VCP/p97-binding molecular-function annotation rather than only generic protein binding? experts: - Alexander Buchberger - Sichen Shao - question: >- Does ANKZF1-mediated RQC limit toxic repeat-associated non-AUG (RAN) translation products in neurons through its canonical tRNA-cleavage/nascent-chain release activity, and is this dependent on catalytic activity rather than a scaffolding role? experts: - Peter K Todd - Sami J Barmada suggested_experiments: - hypothesis: >- ANKZF1's mitochondrial stress phenotype is separable from direct mitochondrial protein catabolism. description: >- In endogenous ANKZF1 knockout cells rescued with wild-type, catalytic-dead, and VIM-mutant ANKZF1, compare turnover of defined mitochondrial protein quality-control substrates with turnover of stalled cytosolic RQC reporters after oxidative stress and translation-stall induction. experiment_type: genetic rescue and quantitative protein-turnover assay - hypothesis: >- ANKZF1 acts on mitochondria-targeted translation products only after they enter the canonical cytosolic RQC substrate state. description: >- Use reporters encoding mitochondrial-targeting sequences followed by defined ribosome-stalling motifs, then assay ANKZF1-dependent tRNA cleavage intermediates, nascent-chain ubiquitination, VCP dependence, and subcellular localization of the stalled complexes. experiment_type: stalled-translation reporter assay