RFWD3

UniProt ID: Q6PCD5
Organism: Homo sapiens
Review Status: COMPLETE
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Gene Description

RFWD3 (RING finger and WD repeat domain-containing protein 3; also RNF201; Fanconi anemia complementation group W, FANCW) is a nuclear RING-type E3 ubiquitin-protein ligase (EC 2.3.2.27). It combines an N-terminal region bearing ATM/ATR phosphorylation sites (Ser46, Ser63), a degenerate RING zinc finger that provides the catalytic ligase activity (active-site Cys315), a coiled-coil, and a C-terminal WD40 Ξ²-propeller that mediates protein-protein interactions. RFWD3 is recruited, through its WD40 domain binding the RPA32 (RPA2) subunit of the single-stranded-DNA-binding replication protein A (RPA) complex, to stalled replication forks and sites of DNA damage. There it polyubiquitinates RPA (all three subunits) and RAD51, promoting their VCP/p97-dependent turnover and timely removal from DNA damage sites so that the RPA-to-RAD51 exchange, homologous recombination and interstrand crosslink (ICL) repair can proceed. RFWD3 also promotes ubiquitination of proteins on single-stranded DNA, driving PCNA ubiquitination and translesion DNA synthesis. It additionally participates in replication-checkpoint signaling (ATR-dependent CHK1 activation) and, as a secondary activity, cooperates with MDM2 to ubiquitinate and stabilize p53 in the late DNA-damage response, contributing to the G1 checkpoint. Biallelic loss-of-function mutations cause Fanconi anemia, and Rfwd3-knockout mice show embryonic lethality, subfertility and reduced lifespan.

Existing Annotations Review

GO Term Evidence Action Reason
GO:0061630 ubiquitin protein ligase activity
IBA
GO_REF:0000033
ACCEPT
Summary: Phylogenetically inferred RING-type ubiquitin ligase activity. This is the core molecular function of RFWD3 and is strongly corroborated by direct biochemistry.
Reason: RFWD3 is a RING-finger E3 ligase (EC 2.3.2.27) with catalytic Cys315; multiple IDA studies demonstrate ligase activity toward RPA, RAD51 and p53. The IBA is at the correct level of specificity and represents the central evolved function.
Supporting Evidence:
PMID:26474068
We demonstrate that the E3 ligase RFWD3 mediates RPA ubiquitination.
GO:0016567 protein ubiquitination
IBA
GO_REF:0000033
ACCEPT
Summary: Phylogenetically inferred involvement in protein ubiquitination, the process carried out by the RFWD3 ligase activity.
Reason: Directly supported: RFWD3 ubiquitinates RPA, RAD51 and p53 and promotes ubiquitination of proteins on ssDNA. Correct and well-supported.
Supporting Evidence:
PMID:33321094
the E3 ubiquitin ligase RFWD3 promotes ubiquitylation of proteins on ssDNA
GO:0005634 nucleus
IBA
GO_REF:0000033
ACCEPT
Summary: Phylogenetically inferred nuclear localization; RFWD3 acts in the nucleus at replication forks and DNA damage sites.
Reason: RFWD3 functions in the nucleus where it ubiquitinates RPA/RAD51 at chromatin; is_active_in nucleus is the correct core location.
Supporting Evidence:
PMID:28691929
impaired relocation of mutant RFWD3 to chromatin
GO:0031297 replication fork processing
IBA
GO_REF:0000033
ACCEPT
Summary: Phylogenetically inferred role at stalled replication forks, consistent with RFWD3 promoting fork restart and repair at stalled forks.
Reason: RFWD3 is recruited to stalled replication forks and is required for fork restart and repair at stalled forks (Elia et al. 2015).
Supporting Evidence:
PMID:26474068
RFWD3 is necessary for replication fork restart, normal repair kinetics during
GO:0090734 site of DNA damage
IBA
GO_REF:0000033
ACCEPT
Summary: Phylogenetically inferred activity at sites of DNA damage; RFWD3 is recruited to and functions at DNA damage sites.
Reason: RFWD3 translocates to tracks of laser micro-irradiation and to MMC-induced foci that co-localize with RPA and gamma-H2AX; this is a core functional location.
Supporting Evidence:
PMID:28575657
translocates to tracks of laser micro-irradiation and co-localizes with the phosphorylated form of histone variant H2AX
GO:0004842 ubiquitin-protein transferase activity
IEA
GO_REF:0000002
ACCEPT
Summary: InterPro-based electronic annotation of ubiquitin-protein transferase activity, a parent/equivalent of the demonstrated RING ligase activity.
Reason: Correct: RFWD3 is a ubiquitin transferase (RING E3). Slightly more general than GO:0061630 but not wrong; the specific ligase activity is captured by the IDA annotations.
GO:0005634 nucleus
IEA
GO_REF:0000120
ACCEPT
Summary: Electronic annotation of nuclear localization, consistent with experimental data.
Reason: Nuclear localization is experimentally established (multiple EXP/IDA annotations); the IEA is correct.
GO:0005737 cytoplasm
IEA
GO_REF:0000044
KEEP AS NON CORE
Summary: Electronic subcellular-location annotation of cytoplasm; RFWD3 is found in the cytoplasm of undamaged cells before nuclear recruitment upon damage.
Reason: A cytoplasmic pool is documented in undamaged cells, but the functional site of RFWD3 is the nucleus/chromatin; cytoplasmic localization is peripheral to its DNA-repair function.
Supporting Evidence:
PMID:21558276
RFWD3 associates with replication protein
GO:0016567 protein ubiquitination
IEA
GO_REF:0000120
ACCEPT
Summary: Electronic annotation of protein ubiquitination, matching experimental evidence.
Reason: RFWD3 ubiquitinates multiple substrates; the process annotation is correct.
GO:0016605 PML body
IEA
GO_REF:0000044
KEEP AS NON CORE
Summary: Electronic subcellular-location annotation of PML nuclear body, from the reported partial association of RFWD3 with PML bodies in undamaged cells.
Reason: RFWD3 is partially associated with PML nuclear bodies in undamaged cells, but this is not the site of its characterized DNA-repair ligase activity.
Supporting Evidence:
PMID:21558276
RFWD3 associates with replication protein
GO:0036297 interstrand cross-link repair
IEA
GO_REF:0000002
ACCEPT
Summary: InterPro-based annotation of interstrand cross-link repair, a core RFWD3 process confirmed experimentally.
Reason: RFWD3-deficient cells are profoundly hypersensitive to ICL-inducing agents; ICL repair is a major RFWD3 function (FANCW).
Supporting Evidence:
PMID:28575657
show profound defects in ICL repair
GO:0061630 ubiquitin protein ligase activity
IEA
GO_REF:0000003
ACCEPT
Summary: EC-mapping-based annotation of ubiquitin ligase activity (EC 2.3.2.27).
Reason: Consistent with the RecName EC 2.3.2.27 and direct biochemical demonstration of RING E3 ligase activity.
GO:0005515 protein binding
IPI
PMID:19549727
Analysis of the human E2 ubiquitin conjugating enzyme protei...
MARK AS OVER ANNOTATED
Summary: IntAct-curated interaction with the E2 conjugating enzyme UBE2N (UBC13) from a systematic E2 interaction-network screen.
Reason: GO:0005515 protein binding is uninformative as a molecular function. The biologically relevant interaction (RFWD3 partnering with the K63-forming E2 UBE2N) is better captured by the ligase activity core function; no specific MF term is warranted from this high-throughput screen.
GO:0005515 protein binding
IPI
PMID:20173098
RFWD3-Mdm2 ubiquitin ligase complex positively regulates p53...
MARK AS OVER ANNOTATED
Summary: IntAct-curated interactions with p53 (TP53) and MDM2 from the RFWD3-MDM2-p53 study.
Reason: Generic protein binding is uninformative; the specific and informative interactions are separately annotated as p53 binding (GO:0002039) and MDM2/MDM4 family protein binding (GO:0097371).
GO:0005515 protein binding
IPI
PMID:24126761
hPrimpol1/CCDC111 is a human DNA primase-polymerase required...
MARK AS OVER ANNOTATED
Summary: IntAct-curated RFWD3-RPA2 (P15927) interaction detected in a PRIMPOL study.
Reason: Uninformative generic binding term; the functionally important RPA2 interaction underlies the core RFWD3 recruitment/ubiquitination function and is captured there.
GO:0005515 protein binding
IPI
PMID:25260751
The MEKK1 PHD ubiquitinates TAB1 to activate MAPKs in respon...
MARK AS OVER ANNOTATED
Summary: IntAct-curated RFWD3-UBE2N (P61088) interaction detected in a MEKK1/TAB1 study.
Reason: Generic protein binding is uninformative; the UBE2N (E2) partnership is subsumed by the ligase activity annotation.
GO:0005515 protein binding
IPI
PMID:28691929
Biallelic mutations in the ubiquitin ligase RFWD3 cause Fanc...
MARK AS OVER ANNOTATED
Summary: IntAct-curated RFWD3-RPA2 (P15927) interaction from the FANCW paper.
Reason: Uninformative generic term; the RPA2 interaction that recruits RFWD3 to chromatin is central and captured by the core function rather than by GO:0005515.
GO:0005515 protein binding
IPI
PMID:40205054
Multimodal cell maps as a foundation for structural and func...
MARK AS OVER ANNOTATED
Summary: IntAct-curated RFWD3-RPA2 (P15927) interaction from a systems-level multimodal cell-mapping study.
Reason: Generic protein binding is uninformative; the RPA2 interaction is already represented functionally elsewhere.
GO:0005654 nucleoplasm
IDA
GO_REF:0000052
ACCEPT
Summary: HPA immunofluorescence localization to the nucleoplasm.
Reason: Consistent with the established nuclear localization; nucleoplasm is an accurate sub-nuclear location.
GO:0005829 cytosol
IDA
GO_REF:0000052
KEEP AS NON CORE
Summary: HPA immunofluorescence localization to the cytosol.
Reason: A cytosolic pool is consistent with cytoplasmic localization in undamaged cells, but is peripheral to the nuclear DNA-repair function.
GO:0005634 nucleus
EXP
PMID:21504906
E3 ligase RFWD3 participates in replication checkpoint contr...
ACCEPT
Summary: Experimental nuclear localization; RFWD3 is recruited to the nucleus/stalled forks upon replication stress.
Reason: Nuclear localization is well established experimentally.
Supporting Evidence:
PMID:21504906
RFWD3 is recruited to stalled replication forks and co-localizes with RPA2 in response to replication stress
GO:0005634 nucleus
EXP
PMID:21558276
RING finger and WD repeat domain 3 (RFWD3) associates with r...
ACCEPT
Summary: Experimental nuclear localization from the RFWD3-RPA association study.
Reason: Consistent with established nuclear localization.
GO:0005634 nucleus
EXP
PMID:28575657
RPA-Mediated Recruitment of the E3 Ligase RFWD3 Is Vital for...
ACCEPT
Summary: Experimental nuclear localization; GFP-RFWD3 forms nuclear foci/tracks at DNA damage sites.
Reason: Nuclear localization at DNA damage sites is directly demonstrated.
Supporting Evidence:
PMID:28575657
translocates to tracks of laser micro-irradiation
GO:0005737 cytoplasm
EXP
PMID:21558276
RING finger and WD repeat domain 3 (RFWD3) associates with r...
KEEP AS NON CORE
Summary: Experimental cytoplasmic localization; RFWD3 is found in the cytoplasm of undamaged cells.
Reason: Real localization in undamaged cells but peripheral to the nuclear repair function.
GO:0005737 cytoplasm
EXP
PMID:28691929
Biallelic mutations in the ubiquitin ligase RFWD3 cause Fanc...
KEEP AS NON CORE
Summary: Experimental cytoplasmic localization reported in the FANCW study.
Reason: Cytoplasmic pool documented; the functionally critical relocation is to chromatin/nucleus.
GO:0036297 interstrand cross-link repair
IDA
PMID:33321094
The ubiquitin ligase RFWD3 is required for translesion DNA s...
ACCEPT
Summary: Direct demonstration of RFWD3's role in the crosslink/lesion-bypass response; RFWD3 promotes ssDNA-protein ubiquitination required for TLS during ICL repair.
Reason: RFWD3 is required for translesion synthesis, an integral gap-filling step of ICL repair; ICL repair involvement is a core function.
Supporting Evidence:
PMID:33321094
TLS across different DNA lesions is drastically impaired
GO:0061630 ubiquitin protein ligase activity
IDA
PMID:20173098
RFWD3-Mdm2 ubiquitin ligase complex positively regulates p53...
ACCEPT
Summary: Direct demonstration of RFWD3 E3 ligase activity toward p53 (in the RFWD3-MDM2 complex).
Reason: In vitro experiments show RFWD3 acts as a p53 E3 ligase; supports the core RING ligase activity. (The p53 process context is secondary.)
Supporting Evidence:
PMID:20173098
RFWD3 is a p53 E3 ubiquitin ligase
GO:0061630 ubiquitin protein ligase activity
IDA
PMID:26474068
RFWD3-Dependent Ubiquitination of RPA Regulates Repair at St...
ACCEPT
Summary: Direct demonstration that RFWD3 ligase activity ubiquitinates RPA.
Reason: Core catalytic function; RFWD3 mediates RPA ubiquitination.
Supporting Evidence:
PMID:26474068
the E3 ligase RFWD3 is responsible for RPA ubiquitination
GO:0061630 ubiquitin protein ligase activity
IDA
PMID:28575657
RPA-Mediated Recruitment of the E3 Ligase RFWD3 Is Vital for...
ACCEPT
Summary: Direct demonstration of RFWD3 ligase activity; the recombinant C315A active-site mutant is inactive in vitro.
Reason: Confirms catalytic RING E3 activity dependent on Cys315; core molecular function.
Supporting Evidence:
PMID:28575657
which renders the recombinant protein inactive in vitro
GO:0061630 ubiquitin protein ligase activity
IDA
PMID:28575658
RFWD3-Mediated Ubiquitination Promotes Timely Removal of Bot...
ACCEPT
Summary: Direct demonstration that RFWD3 polyubiquitinates both RPA and RAD51 in vitro and in vivo.
Reason: Core catalytic function toward the physiological substrates RPA and RAD51.
Supporting Evidence:
PMID:28575658
RFWD3 polyubiquitinates both RPA and RAD51 in vitro and in vivo
GO:0000724 double-strand break repair via homologous recombination
IMP
PMID:28691929
Biallelic mutations in the ubiquitin ligase RFWD3 cause Fanc...
ACCEPT
Summary: Loss-of-function (patient/engineered mutations) disrupts homologous recombination, establishing RFWD3's role in HR.
Reason: HR was disrupted in RFWD3-mutant cells; a core process for RFWD3 (FANCW).
Supporting Evidence:
PMID:28691929
HR was disrupted in RFWD3-mutant cells
GO:0005634 nucleus
IMP
PMID:28691929
Biallelic mutations in the ubiquitin ligase RFWD3 cause Fanc...
ACCEPT
Summary: Mutation-based evidence that RFWD3 relocation to the nucleus/chromatin is required for function.
Reason: Nuclear/chromatin relocation is impaired by FA mutations, underscoring the nucleus as the functional compartment.
Supporting Evidence:
PMID:28691929
impaired relocation of mutant RFWD3 to chromatin
GO:0000724 double-strand break repair via homologous recombination
IDA
PMID:26474068
RFWD3-Dependent Ubiquitination of RPA Regulates Repair at St...
ACCEPT
Summary: RFWD3 is required for homologous recombination at stalled replication forks.
Reason: Core process; RFWD3 stimulates HR at stalled forks.
Supporting Evidence:
PMID:26474068
homologous recombination (HR) at stalled replication forks
GO:0000724 double-strand break repair via homologous recombination
IDA
PMID:28575657
RPA-Mediated Recruitment of the E3 Ligase RFWD3 Is Vital for...
ACCEPT
Summary: RFWD3-deficient cells show HR defects (PARP-inhibitor hypersensitivity, persistent RPA in RAD51 foci).
Reason: Supports the core HR function; RFWD3 controls RPA/RAD51 dynamics required for HR.
Supporting Evidence:
PMID:28575657
unloading of RPA from sites of ICL induction is perturbed in RFWD3-deficient cells
GO:0000724 double-strand break repair via homologous recombination
IDA
PMID:28575658
RFWD3-Mediated Ubiquitination Promotes Timely Removal of Bot...
ACCEPT
Summary: RFWD3-mediated removal of RPA and RAD51 facilitates late-phase homologous recombination.
Reason: Core HR process, mechanistically defined via RPA/RAD51 turnover.
Supporting Evidence:
PMID:28575658
crucial for progression to the late-phase HR and suppression of the FA phenotype
GO:0005515 protein binding
IPI
PMID:28575657
RPA-Mediated Recruitment of the E3 Ligase RFWD3 Is Vital for...
MARK AS OVER ANNOTATED
Summary: IntAct-curated interactions with RPA2 (P15927) and RAD51 (Q06609), the physiological RFWD3 substrates/partners.
Reason: Generic protein binding is uninformative as an MF; the functionally meaningful RPA2 and RAD51 interactions underpin the core ligase/HR function and are captured there.
GO:0005515 protein binding
IPI
PMID:28575658
RFWD3-Mediated Ubiquitination Promotes Timely Removal of Bot...
MARK AS OVER ANNOTATED
Summary: IntAct-curated RFWD3-RPA2 (P15927) interaction.
Reason: Uninformative generic binding term; the RPA2 interaction is functionally represented by the core function.
GO:0006974 DNA damage response
IDA
PMID:26474068
RFWD3-Dependent Ubiquitination of RPA Regulates Repair at St...
ACCEPT
Summary: RFWD3 acts within the DNA damage response (RPA ubiquitination upon replication stress).
Reason: Accurate high-level process; RFWD3's ligase activity is part of the DDR. More specific processes (HR, ICL repair, fork processing) are also annotated.
Supporting Evidence:
PMID:26474068
to profile ubiquitination in the DNA damage response (DDR)
GO:0006974 DNA damage response
IDA
PMID:28575657
RPA-Mediated Recruitment of the E3 Ligase RFWD3 Is Vital for...
ACCEPT
Summary: RFWD3 recruitment and action at DNA damage sites within the DDR.
Reason: Correct high-level process annotation, consistent with damage-site recruitment.
Supporting Evidence:
PMID:28575657
translocates to tracks of laser micro-irradiation
GO:0006974 DNA damage response
IDA
PMID:28575658
RFWD3-Mediated Ubiquitination Promotes Timely Removal of Bot...
ACCEPT
Summary: RFWD3 acts in the DDR via ATR/ATM-regulated ubiquitination of RPA and RAD51.
Reason: Correct high-level DDR process; RFWD3 activity depends on ATR/ATM phosphorylation.
Supporting Evidence:
PMID:28575658
Phosphorylation by ATR and ATM kinases is required for this activity in vivo
GO:0016567 protein ubiquitination
IDA
PMID:26474068
RFWD3-Dependent Ubiquitination of RPA Regulates Repair at St...
ACCEPT
Summary: Direct evidence of RFWD3-mediated RPA ubiquitination.
Reason: Core process; RFWD3 ubiquitinates RPA on chromatin.
Supporting Evidence:
PMID:26474068
the E3 ligase RFWD3 is responsible for RPA ubiquitination
GO:0016567 protein ubiquitination
IDA
PMID:28575657
RPA-Mediated Recruitment of the E3 Ligase RFWD3 Is Vital for...
ACCEPT
Summary: RFWD3 ubiquitinates RPA at sites of ICL-induced fork stalling.
Reason: Core process; RFWD3 ubiquitylates RPA.
Supporting Evidence:
PMID:28575657
interacts with and ubiquitylates replication protein A (RPA)
GO:0016567 protein ubiquitination
IDA
PMID:28575658
RFWD3-Mediated Ubiquitination Promotes Timely Removal of Bot...
ACCEPT
Summary: RFWD3 polyubiquitinates RPA and RAD51.
Reason: Core process, defined toward physiological substrates.
Supporting Evidence:
PMID:28575658
RFWD3 polyubiquitinates both RPA and RAD51 in vitro and in vivo
GO:0031297 replication fork processing
IDA
PMID:26474068
RFWD3-Dependent Ubiquitination of RPA Regulates Repair at St...
ACCEPT
Summary: RFWD3 promotes restart and remodeling of stalled replication forks and supports normal fork progression.
Reason: Direct evidence that RFWD3 is necessary for fork restart and repair at stalled forks (Elia et al. 2015); further corroborated by its role in normal fork progression via PCNA binding (Lin et al. 2018) and in ZRANB3-dependent fork remodeling/reversal (Moore et al. 2023).
Supporting Evidence:
PMID:26474068
we found that depletion of RFWD3 decreased fork restart
PMID:30530694
in unperturbed human cells, RFWD3 localizes at replication forks and associates with proliferating cell nuclear antigen (PCNA) via its PCNA-interacting protein (PIP) motif
PMID:37036693
RFWD3 stimulates fork remodeling in a ZRANB3-epistatic manner
GO:0036297 interstrand cross-link repair
IMP
PMID:28575657
RPA-Mediated Recruitment of the E3 Ligase RFWD3 Is Vital for...
ACCEPT
Summary: RFWD3-hypomorphic cells are profoundly hypersensitive to ICL-inducing agents, demonstrating an ICL-repair role.
Reason: Loss-of-function evidence establishes ICL repair as a major RFWD3 function (core).
Supporting Evidence:
PMID:28575657
argues strongly that ICL repair is a major function of RFWD3
GO:0036297 interstrand cross-link repair
IMP
PMID:28575658
RFWD3-Mediated Ubiquitination Promotes Timely Removal of Bot...
ACCEPT
Summary: RFWD3 inactivation impairs ICL repair; suppression of the FA phenotype depends on RFWD3-mediated RPA/RAD51 removal.
Reason: Core ICL-repair process consistent with FANCW identity.
Supporting Evidence:
PMID:28575658
suppression of the FA phenotype
GO:0090734 site of DNA damage
IDA
PMID:28575657
RPA-Mediated Recruitment of the E3 Ligase RFWD3 Is Vital for...
ACCEPT
Summary: RFWD3 localizes to sites of ICL/DNA damage (laser tracks, MMC foci).
Reason: Core functional location, directly imaged.
Supporting Evidence:
PMID:28575657
translocates to tracks of laser micro-irradiation
GO:0090734 site of DNA damage
IDA
PMID:28575658
RFWD3-Mediated Ubiquitination Promotes Timely Removal of Bot...
ACCEPT
Summary: RFWD3 acts at DNA damage sites to remove RPA and RAD51.
Reason: Consistent with damage-site localization and function; core functional location.
Supporting Evidence:
PMID:28575658
timely removal of RPA and RAD51 from DNA damage sites
GO:0005515 protein binding
IPI
PMID:21504906
E3 ligase RFWD3 participates in replication checkpoint contr...
MARK AS OVER ANNOTATED
Summary: IntAct-curated direct RFWD3-RPA2 (P15927) interaction, shown with purified proteins.
Reason: Generic protein binding is uninformative; the direct RPA2 interaction that recruits RFWD3 to stalled forks is central and represented by the core function.
GO:0035861 site of double-strand break
IDA
PMID:21504906
E3 ligase RFWD3 participates in replication checkpoint contr...
ACCEPT
Summary: RFWD3 co-localizes with RPA2 at stalled forks/damage sites in response to replication stress.
Reason: Accurate colocalization annotation consistent with recruitment to damage sites; complements the site of DNA damage annotations.
Supporting Evidence:
PMID:21504906
RFWD3 is recruited to stalled replication forks and co-localizes with RPA2 in response to replication stress
GO:2000001 regulation of DNA damage checkpoint
IMP
PMID:21504906
E3 ligase RFWD3 participates in replication checkpoint contr...
KEEP AS NON CORE
Summary: RFWD3 depletion decreases ATR-dependent CHK1 activation after replication stress, implicating it in replication-checkpoint control.
Reason: A genuine but secondary/context-dependent role; the effect on CHK1 activation is cell-type dependent and is distinct from the core RPA/RAD51-ubiquitination repair function.
Supporting Evidence:
PMID:21504906
RFWD3 is important for ATR-dependent Chk1 activation in response to replication stress
GO:0002039 p53 binding
IPI
PMID:20173098
RFWD3-Mdm2 ubiquitin ligase complex positively regulates p53...
KEEP AS NON CORE
Summary: Direct interaction with p53 (TP53) within the RFWD3-MDM2-p53 complex that regulates p53 stability.
Reason: An informative, specific binding (unlike generic protein binding), but it underlies the secondary p53-stabilization role rather than the core DNA-repair ligase function.
Supporting Evidence:
PMID:20173098
forms a complex with Mdm2 and p53 to synergistically ubiquitinate p53
GO:0005634 nucleus
IDA
PMID:20173098
RFWD3-Mdm2 ubiquitin ligase complex positively regulates p53...
ACCEPT
Summary: Experimental nuclear localization in the p53-regulation study.
Reason: Consistent with established nuclear localization.
GO:0010212 response to ionizing radiation
IDA
PMID:20173098
RFWD3-Mdm2 ubiquitin ligase complex positively regulates p53...
KEEP AS NON CORE
Summary: RFWD3 is phosphorylated and stabilizes p53 in the late response to ionizing radiation.
Reason: A real damage-response context, but tied to the secondary p53 role; the core RFWD3 function is better described by HR/ICL-repair processes.
Supporting Evidence:
PMID:20173098
p53 stabilization in the late phase after ionizing radiation correlates with active ubiquitination
GO:0016567 protein ubiquitination
IDA
PMID:20173098
RFWD3-Mdm2 ubiquitin ligase complex positively regulates p53...
ACCEPT
Summary: RFWD3 ubiquitinates p53 (with MDM2) in the late DNA-damage response.
Reason: A genuine protein-ubiquitination event catalyzed by RFWD3; correct process term, here in the secondary p53 context.
Supporting Evidence:
PMID:20173098
forms a complex with Mdm2 and p53 to synergistically ubiquitinate p53
GO:0031571 mitotic G1 DNA damage checkpoint signaling
IMP
PMID:20173098
RFWD3-Mdm2 ubiquitin ligase complex positively regulates p53...
KEEP AS NON CORE
Summary: RFWD3 positively regulates p53 stability when the G1 cell-cycle checkpoint is activated.
Reason: Secondary function via p53/MDM2; distinct from the core replication-associated repair activity and supported by a single group.
Supporting Evidence:
PMID:20173098
a positive regulator of p53 stability when the G(1) cell cycle checkpoint is activated
GO:0097371 MDM2/MDM4 family protein binding
IPI
PMID:20173098
RFWD3-Mdm2 ubiquitin ligase complex positively regulates p53...
KEEP AS NON CORE
Summary: Direct interaction with MDM2 in the p53-regulatory RFWD3-MDM2 complex.
Reason: Informative specific binding supporting the secondary p53-stabilization role, not the core DNA-repair ligase function.
Supporting Evidence:
PMID:20173098
RFWD3 (RNF201/FLJ10520) forms a complex with Mdm2 and p53

Core Functions

RING-type E3 ubiquitin-protein ligase that is recruited via its C-terminal WD40 domain to RPA-coated single-stranded DNA at stalled replication forks and sites of DNA damage, where (following ATM/ATR phosphorylation) it polyubiquitinates the RPA complex and RAD51 to drive their VCP/p97-dependent turnover and timely removal, enabling homologous recombination and interstrand crosslink repair to progress.

Supporting Evidence:

References

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Deep Research

Affinage

(RFWD3-deep-research-affinage.md)

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πŸ“š Additional Documentation

Notes

(RFWD3-notes.md)

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