RNF25 (RING finger protein 25, originally identified as AO7) is a cytoplasmic RING-H2 E3 ubiquitin-protein ligase that, with an N-terminal RWD domain for E2 recognition, functions in ribosome-associated quality control. It is a core component of the RNF14-RNF25 translation quality control pathway that operates when ribosomes stall and collide during translation. RNF25 catalyzes ubiquitination of the ribosomal protein RPS27A/eS31 in response to ribosome collisions, providing a signal that activates the partner ligase RNF14, and it ubiquitinates additional ribosomal proteins and stalled release factor ETF1/eRF1, marking translation factors on stalled ribosomes for degradation. RNF25 also assembles atypical K6-linked ubiquitin chains that flag formaldehyde-induced RNA-protein crosslinks for translation-coupled resolution. It works with the E2 enzyme UBE2D2 (UbcH5B). Independently of the stalled-ribosome response, RNF25 was originally characterized as a regulator that supports NF-kappaB (RELA/p65)-mediated transcription and can target substrates such as NKD2 for degradation.
| GO Term | Evidence | Action | Reason |
|---|---|---|---|
| GO:0072344 rescue of stalled cytosolic ribosome | IBA GO_REF:0000033 | ACCEPT | Summary: RNF25 acts on stalled/collided ribosomes in the RNF14-RNF25 quality control pathway. IBA inference is corroborated by direct experimental evidence. Reason: Core process; RNF25 ubiquitinates ribosomal proteins and translation factors on stalled ribosomes as part of ribosome-associated quality control. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt E3 ubiquitin-protein ligase that plays a key role in the RNF14-RNF25 translation quality control pathway |
| GO:0006511 ubiquitin-dependent protein catabolic process | IBA GO_REF:0000033 | ACCEPT | Summary: RNF25 ubiquitinates translation factors and ribosomal proteins on stalled ribosomes, targeting them for degradation. Core biological process. Reason: Directly supported; RNF25 promotes ubiquitin-dependent degradation of stalled translation factors (e.g. eEF1A, eRF1). Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt promotes ubiquitination and degradation of translation factors on stalled ribosomes |
| GO:0061630 ubiquitin protein ligase activity | IBA GO_REF:0000033 | ACCEPT | Summary: RNF25 is a RING E3 ubiquitin ligase. This is its core molecular function. Reason: Defining molecular function, supported by phylogenetic inference and direct biochemistry. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt E3 ubiquitin-protein ligase that plays a key role in the RNF14-RNF25 translation quality control pathway |
| GO:0005634 nucleus | IBA GO_REF:0000033 | KEEP AS NON CORE | Summary: A nuclear pool of RNF25 is reported (from its AO7/NF-kB role), but its principal quality-control function is cytoplasmic/ribosome-associated. Reason: Nuclear localization is documented but peripheral to the core cytoplasmic RQC function. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt may also stimulate transcription mediated by NF-kappa-B via its interaction with RELA/p65 |
| GO:0005737 cytoplasm | IEA GO_REF:0000044 | ACCEPT | Summary: Cytoplasmic localization, consistent with RNF25's ribosome-associated quality-control function. Reason: Correct compartment for the core RQC function. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt SUBCELLULAR LOCATION: Cytoplasm |
| GO:0061630 ubiquitin protein ligase activity | IEA GO_REF:0000120 | ACCEPT | Summary: Electronic transfer of ubiquitin ligase activity, consistent with direct evidence. Reason: Correct core molecular function. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt E3 ubiquitin-protein ligase that plays a key role in the RNF14-RNF25 translation quality control pathway |
| GO:0005515 protein binding | IPI PMID:18757723 EGF receptor-independent action of TGF-alpha protects Naked2... | KEEP AS NON CORE | Summary: High-throughput interaction. Bare protein binding is uninformative. Reason: Records a physical interaction but the generic term adds nothing to RNF25's E3 ligase function. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt E3 ubiquitin-protein ligase that plays a key role in the RNF14-RNF25 translation quality control pathway |
| GO:0005515 protein binding | IPI PMID:19549727 Analysis of the human E2 ubiquitin conjugating enzyme protei... | KEEP AS NON CORE | Summary: E2 ubiquitin-conjugating enzyme interaction network capturing RNF25-E2 (UbcH5/UBE2D) binding. Relevant to catalysis but recorded as bare protein binding. Reason: The E2 interaction underlies RNF25's ligase activity, but the generic protein binding term is uninformative; the catalytic role is captured by the ligase-activity term. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt Interacts with UBE2D2 |
| GO:0005515 protein binding | IPI PMID:19690564 A comprehensive framework of E2-RING E3 interactions of the ... | KEEP AS NON CORE | Summary: E2-RING E3 interaction framework capturing RNF25-E2 binding. Bare protein binding. Reason: Reflects E2 engagement underlying catalysis; generic term is non-core. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt Interacts with UBE2D2 |
| GO:0005515 protein binding | IPI PMID:28514442 Architecture of the human interactome defines protein commun... | KEEP AS NON CORE | Summary: Interactome screen capturing RNF25 interactions. Bare protein binding. Reason: Generic term; uninformative for core function. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt E3 ubiquitin-protein ligase that plays a key role in the RNF14-RNF25 translation quality control pathway |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | KEEP AS NON CORE | Summary: Binary interactome (e.g. NKD2, a substrate; KCTD21). Bare protein binding. Reason: Records interactions including a substrate (NKD2), but the generic term is uninformative. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt Q969F2: NKD2 |
| GO:0005515 protein binding | IPI PMID:32814053 Interactome Mapping Provides a Network of Neurodegenerative ... | KEEP AS NON CORE | Summary: Neurodegeneration interactome screen. Bare protein binding with high-throughput partners unrelated to the core function. Reason: Isolated high-throughput interactions; generic term uninformative and not part of the core function. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt E3 ubiquitin-protein ligase that plays a key role in the RNF14-RNF25 translation quality control pathway |
| GO:0005515 protein binding | IPI PMID:33961781 Dual proteome-scale networks reveal cell-specific remodeling... | KEEP AS NON CORE | Summary: BioPlex affinity-purification interactome capturing RNF25 interactions. Bare protein binding. Reason: Generic term; uninformative for core function. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt E3 ubiquitin-protein ligase that plays a key role in the RNF14-RNF25 translation quality control pathway |
| GO:0005515 protein binding | IPI PMID:40205054 Multimodal cell maps as a foundation for structural and func... | KEEP AS NON CORE | Summary: Multimodal cell-maps interactome capturing RNF25 interactions. Bare protein binding. Reason: Generic term; uninformative for core function. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt E3 ubiquitin-protein ligase that plays a key role in the RNF14-RNF25 translation quality control pathway |
| GO:0004842 ubiquitin-protein transferase activity | IEA GO_REF:0000107 | ACCEPT | Summary: Ubiquitin-protein transferase activity, equivalent to RNF25's E3 ligase function. Reason: Correct molecular function (the EC 2.3.2.27 ligase reaction). Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt E3 ubiquitin-protein ligase that plays a key role in the RNF14-RNF25 translation quality control pathway |
| GO:0016567 protein ubiquitination | IEA GO_REF:0000120 | ACCEPT | Summary: RNF25 ubiquitinates substrate proteins; protein ubiquitination is the process it catalyzes. Reason: Directly supported core process. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt promotes ubiquitination and degradation of translation factors on stalled ribosomes |
| GO:0005737 cytoplasm | ISS GO_REF:0000024 | ACCEPT | Summary: Sequence-similarity transfer of cytoplasmic localization, consistent with direct evidence. Reason: Correct compartment for the core function. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt SUBCELLULAR LOCATION: Cytoplasm |
| GO:0022626 cytosolic ribosome | IDA PMID:36638793 An E3 ligase network engages GCN1 to promote the degradation... | ACCEPT | Summary: RNF25 acts at the cytosolic ribosome, ubiquitinating ribosomal proteins on stalled/collided ribosomes. Reason: Directly supported site of action for the RQC function. Supporting Evidence: PMID:36638793 RNF25-dependent ubiquitination of the ribosomal protein |
| GO:0061630 ubiquitin protein ligase activity | IDA PMID:36638793 An E3 ligase network engages GCN1 to promote the degradation... | ACCEPT | Summary: Direct demonstration of RNF25 ubiquitin ligase activity in the RNF14-RNF25 pathway, ubiquitinating RPS27A and translation factors on stalled ribosomes. Reason: Directly demonstrated core molecular function in the GCN1-engaging E3 network. Supporting Evidence: PMID:36638793 RNF25-dependent ubiquitination of the ribosomal protein |
| GO:0072344 rescue of stalled cytosolic ribosome | IDA PMID:36638793 An E3 ligase network engages GCN1 to promote the degradation... | ACCEPT | Summary: RNF25 acts on stalled/collided ribosomes, ubiquitinating RPS27A to activate RNF14 and promote degradation of stalled translation factors. Reason: Directly demonstrated involvement in the stalled-ribosome quality-control response. Supporting Evidence: PMID:36638793 RNF25-dependent ubiquitination of the ribosomal protein |
| GO:0061630 ubiquitin protein ligase activity | IDA PMID:37951215 K6-linked ubiquitylation marks formaldehyde-induced RNA-prot... | ACCEPT | Summary: Direct demonstration of RNF25 E3 ligase activity in marking RNA-protein crosslinks with ubiquitin. Reason: Directly demonstrated core molecular function. Supporting Evidence: PMID:37951215 K6-linked ubiquitylation marks formaldehyde-induced RNA-protein crosslinks |
| GO:0160127 protein-RNA covalent cross-linking repair | IDA PMID:37951215 K6-linked ubiquitylation marks formaldehyde-induced RNA-prot... | ACCEPT | Summary: RNF25 (with RNF14) assembles K6-linked ubiquitin chains that flag RNA-protein crosslinks for translation-coupled resolution. Reason: Directly demonstrated role in resolving covalent RNA-protein crosslinks. Supporting Evidence: PMID:37951215 K6-linked ubiquitylation marks formaldehyde-induced RNA-protein crosslinks |
| GO:0061630 ubiquitin protein ligase activity | IDA PMID:37651229 Drug-induced eRF1 degradation promotes readthrough and revea... | ACCEPT | Summary: Direct demonstration of RNF25 ligase activity in drug-induced eRF1 degradation that promotes readthrough. Reason: Directly demonstrated core molecular function. Supporting Evidence: PMID:37651229 E3 ubiquitin ligases RNF14 and RNF25 |
| GO:0072344 rescue of stalled cytosolic ribosome | IDA PMID:37651229 Drug-induced eRF1 degradation promotes readthrough and revea... | ACCEPT | Summary: RNF25 (with RNF14) acts on stalled/collided ribosomes following eRF1 trapping to clear stalled translation factors. Reason: Directly demonstrated involvement in resolving stalled ribosomes. Supporting Evidence: PMID:37651229 E3 ubiquitin ligases RNF14 and RNF25 |
| GO:0072344 rescue of stalled cytosolic ribosome | IDA PMID:37951216 RNF14-dependent atypical ubiquitylation promotes translation... | ACCEPT | Summary: RNF25 (with RNF14) acts in translation-coupled resolution of RNA-protein crosslinks at collided ribosomes. Reason: Directly demonstrated involvement in stalled-ribosome resolution. Supporting Evidence: PMID:37951216 RNF14-dependent atypical ubiquitylation promotes translation-coupled resolution |
| GO:0061630 ubiquitin protein ligase activity | IDA PMID:26475854 Insights into Ubiquitination from the Unique Clamp-like Bind... | ACCEPT | Summary: Structural/biochemical study of the AO7 (RNF25) RING-E2 (UbcH5B) interaction demonstrating its ubiquitin ligase activity. Reason: Directly demonstrated RING E3 ligase activity with its E2 partner. Supporting Evidence: PMID:26475854 Insights into Ubiquitination from the Unique Clamp-like Binding of the RING E3 AO7 to the E2 UbcH5B |
| GO:0006511 ubiquitin-dependent protein catabolic process | IDA PMID:27863242 Decoding Mammalian Ribosome-mRNA States by Translational GTP... | ACCEPT | Summary: RNF25-dependent ubiquitination targets factors on stalled ribosomes for degradation. Reason: Directly supported core process. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt promotes ubiquitination and degradation of translation factors on stalled ribosomes |
| GO:0022626 cytosolic ribosome | IDA PMID:27863242 Decoding Mammalian Ribosome-mRNA States by Translational GTP... | ACCEPT | Summary: RNF25 acts at the cytosolic ribosome in ribosome-associated quality control. Reason: Directly supported site of action. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt Catalyzes ubiquitination of RPS27A in response to ribosome collisions |
| GO:0061630 ubiquitin protein ligase activity | IDA PMID:27863242 Decoding Mammalian Ribosome-mRNA States by Translational GTP... | ACCEPT | Summary: Direct demonstration of RNF25 ubiquitin ligase activity in the RQC context. Reason: Directly demonstrated core molecular function. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt E3 ubiquitin-protein ligase that plays a key role in the RNF14-RNF25 translation quality control pathway |
| GO:0072344 rescue of stalled cytosolic ribosome | IDA PMID:27863242 Decoding Mammalian Ribosome-mRNA States by Translational GTP... | ACCEPT | Summary: RNF25 acts on stalled/collided ribosomes in ribosome-associated quality control. Reason: Directly supported core process. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt Catalyzes ubiquitination of RPS27A in response to ribosome collisions |
| GO:0085020 protein K6-linked ubiquitination | IDA PMID:27863242 Decoding Mammalian Ribosome-mRNA States by Translational GTP... | ACCEPT | Summary: RNF25 assembles atypical K6-linked ubiquitin chains, a distinctive feature of its RQC signaling. Reason: Directly demonstrated; K6-linked ubiquitination marks RNA-protein crosslinks and stalled-ribosome substrates. Supporting Evidence: PMID:37951215 K6-linked ubiquitylation marks formaldehyde-induced RNA-protein crosslinks |
| GO:0004842 ubiquitin-protein transferase activity | ISS GO_REF:0000024 | ACCEPT | Summary: Sequence-similarity transfer of ubiquitin-protein transferase activity, consistent with direct evidence. Reason: Correct core molecular function. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt E3 ubiquitin-protein ligase that plays a key role in the RNF14-RNF25 translation quality control pathway |
| GO:0005634 nucleus | IDA PMID:12748188 RING finger protein AO7 supports NF-kappaB-mediated transcri... | KEEP AS NON CORE | Summary: Direct nuclear localization reported in the original AO7/NF-kB study. Reason: Genuine nuclear pool linked to the NF-kB role; peripheral to the core cytoplasmic RQC function. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt may also stimulate transcription mediated by NF-kappa-B via its interaction with RELA/p65 |
| GO:0005829 cytosol | IDA PMID:12748188 RING finger protein AO7 supports NF-kappaB-mediated transcri... | ACCEPT | Summary: Direct cytosolic localization, consistent with the core ribosome-associated function. Reason: Correct localization for the cytoplasmic RQC function. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt SUBCELLULAR LOCATION: Cytoplasm |
| GO:0016567 protein ubiquitination | ISS GO_REF:0000024 | ACCEPT | Summary: RNF25 catalyzes protein ubiquitination. Reason: Correct core process. Supporting Evidence: file:human/RNF25/RNF25-uniprot.txt promotes ubiquitination and degradation of translation factors on stalled ribosomes |
| GO:0051059 NF-kappaB binding | IPI PMID:12748188 RING finger protein AO7 supports NF-kappaB-mediated transcri... | KEEP AS NON CORE | Summary: RNF25 (AO7) binds RELA/p65 and supports NF-kappaB-mediated transcription, a function distinct from its ribosome-associated quality-control role. Reason: Documented physical interaction underlying the NF-kB moonlighting role; retained as non-core relative to the E3-ligase RQC function. Supporting Evidence: PMID:12748188 RING finger protein AO7 supports NF-kappaB-mediated transcription |
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Download this section (compressed HTML)Q: What determines the substrate hierarchy in RNF25-mediated ubiquitination (RPS27A vs eRF1 vs other ribosomal proteins) and how does RPS27A ubiquitination activate RNF14?
Q: How is RNF25's RQC E3-ligase role partitioned from its older AO7/NF-kappaB transcriptional role, and are these mediated by distinct pools or stimuli?
Experiment: Site-specific ubiquitin-proteomics (diGly) in RNF25-knockout versus wild-type cells under collision-inducing stress to define the endogenous RNF25 substrate set and ubiquitin-chain linkage types.
Experiment: Reconstituted ubiquitination assays with RNF25, RNF14, UBE2D2 and collided ribosomes to dissect the order of RPS27A ubiquitination and RNF14 activation.
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