Affinage

CNOT4

CCR4-NOT transcription complex subunit 4 · UniProt O95628

Length
575 aa
Mass
63.5 kDa
Annotated
2026-06-09
39 papers in source corpus 33 papers cited in narrative 33 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 7/7 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

CNOT4 (yeast Not4) is a RING-finger E3 ubiquitin ligase that bridges ubiquitin-dependent protein quality control, translation, and the CCR4-NOT mRNA-degradation machinery (PMID:7926748, PMID:15062086). Its N-terminal RING domain coordinates two zinc ions in a cross-brace arrangement and selects UbcH5B as its cognate ubiquitin-conjugating enzyme, an E2/E3 interface structurally distinct from the c-Cbl/UbcH7 paradigm (PMID:11087754, PMID:15062086). CNOT4 was first defined as a peripheral subunit of the CCR4-NOT complex, associating via the C-terminal region of NOT1 in yeast and, in metazoans, through a conserved CAF40-binding motif whose interface is shared with Roquin and Bag-of-marbles (PMID:10490603, PMID:30692204); in human cells it does not stably co-purify with the complex but associates in proximity-labeling and, when depleted, accelerates global mRNA decay opposite to CNOT1, while in mouse germ cells it acts as an mRNA adaptor delivering transcripts to CNOT7 for deadenylation (PMID:34026442, PMID:41161383). As an autonomous ligase CNOT4 ubiquitinates a wide substrate range with linkage- and outcome-specific consequences: K48-linked polyubiquitination and proteasomal degradation of stress-induced cyclin C, PAF1, and YAP (PMID:22421358, PMID:25933433, PMID:37802875); monoubiquitination of the ribosomal protein Rps7A and of influenza nucleoprotein NP (PMID:22243599, PMID:28536288); and K63-linked ubiquitination of ABCE1 to signal mitophagy and of ZNF598 during ribosome quality control (PMID:29861391, PMID:38388640). CNOT4 associates with translating ribosomes, where it shapes elongation dynamics and A-site dwelling at non-optimal codons and couples ribosome pausing to co-translational protein quality control and degradation (PMID:25971775, PMID:34469733, PMID:36803582). It also governs proteasome homeostasis, maintaining proteasome integrity through Ecm29 and acting as an assembly checkpoint by ubiquitinating Rpt5 in competition with assembly chaperones (PMID:21321079, PMID:30530678). CNOT4 abundance is itself controlled by TRIM21/TNKS1BP1-mediated K48/K6 ubiquitination, and its degradation of PAF1 is antagonized by the deubiquitinase USP4 (PMID:39019859, PMID:41857610).

Mechanistic history

Synthesis pass · year-by-year structured walk · 18 steps
  1. 1994 Medium

    Established NOT4 as a physical and functional member of a nuclear NOT complex repressing RNA Pol II transcription, anchoring it to a multiprotein regulatory machine before any enzymatic role was known.

    Evidence Two-hybrid, co-fractionation and suppressor genetics in yeast

    PMID:7926748

    Open questions at the time
    • No biochemical activity assigned to NOT4 itself
    • Architecture within the complex undefined
  2. 1999 Medium

    Mapped NOT4 to a peripheral position on the CCR4-CAF1-NOT1 core, showing it is dispensable for core integrity and distinguishing it from constitutive subunits.

    Evidence Co-IP, two-hybrid and deletion mapping in the yeast CCR4-NOT complex

    PMID:10490603

    Open questions at the time
    • Functional consequence of peripheral association unclear
    • Does not address metazoan binding mode
  3. 2000 High

    Solved the C4C4 RING-finger structure and cross-brace zinc coordination, providing the structural basis for a catalytic identity.

    Evidence Heteronuclear NMR with 113Cd substitution

    PMID:11087754

    Open questions at the time
    • No E2 partner identified
    • No substrate or ligase activity demonstrated
  4. 2004 High

    Defined UbcH5B as the cognate E2 and established the molecular basis of E2/E3 specificity, confirming CNOT4 as a bona fide E3 ligase.

    Evidence NMR chemical-shift mapping, HADDOCK docking and mutagenesis

    PMID:15062086

    Open questions at the time
    • Physiological substrates not yet known
    • Linkage type not determined
  5. 2010 Medium

    Linked Not4 ligase activity with Ubc4 to in vivo turnover of a DNA polymerase subunit, the first cellular substrate context.

    Evidence Yeast deletion genetics, stability assays and HU-sensitivity phenotypes

    PMID:20660159

    Open questions at the time
    • Direct ubiquitination of Cdc17 not reconstituted
    • Linkage type unresolved
  6. 2011 High

    Revealed a role in proteasome homeostasis, with Not4 maintaining proteasome integrity through Ecm29.

    Evidence Proteasome purification, Co-IP, in vivo ubiquitination and deletion genetics in yeast

    PMID:21321079

    Open questions at the time
    • Mechanism by which Not4 prevents aberrant RP-CP complexes incomplete
    • Direct Ecm29 ubiquitination not fully defined
  7. 2012 High

    Identified ribosome-associated and stress-responsive substrates (Rps7A monoubiquitination; K48 degradation of cyclin C; Yap1 turnover), establishing CNOT4 as a multi-substrate ligase coupled to translation and oxidative stress.

    Evidence In vitro and in vivo ubiquitination, polysome fractionation, localization and pathway epistasis in yeast

    PMID:22243599 PMID:22421358 PMID:22707721

    Open questions at the time
    • Functional outcome of Rps7A monoubiquitination on the ribosome not resolved
    • How substrate selection is achieved unknown
  8. 2014 Medium

    Connected Not4 to co-translational quality control, showing it represses translation of stall-prone transcripts and clears aberrant proteins independently of the Ccr4 deadenylase module.

    Evidence Polysome fractionation, aggregation assays, pulse-chase and genetic epistasis in yeast

    PMID:24465968 PMID:25971775

    Open questions at the time
    • Direct ribosome-bound substrates of repression unidentified
    • Mechanistic coupling of ligase activity to translational repression unclear
  9. 2015 Medium

    Extended substrate range to human PAF1, demonstrating K48-linked polyubiquitination and proteasomal degradation requiring nuclear but not chromatin localization.

    Evidence Co-IP, ubiquitination assay and NLS-mutant analysis in human cells

    PMID:25933433

    Open questions at the time
    • Upstream signal controlling PAF1 degradation unknown
    • Physiological context not yet defined
  10. 2017 High

    Showed CNOT4 ubiquitinates a viral substrate, monoubiquitinating influenza NP in its RNA-binding groove to promote viral replication without altering NP stability.

    Evidence Co-IP, in vitro ubiquitination, MS site mapping and arginine-substitution mutagenesis

    PMID:28536288

    Open questions at the time
    • Host benefit/cost of this activity unresolved
    • Whether host substrates share this monoubiquitination mode unknown
  11. 2018 High

    Defined linkage-specific signaling roles: K63 ubiquitination of ABCE1 to trigger mitophagy and Rpt5 ubiquitination as a proteasome assembly checkpoint that blocks Rpn1/Ubp6 incorporation.

    Evidence In vitro reconstitution, structural modeling, Drosophila PINK1 epistasis and ribosome fractionation

    PMID:29861391 PMID:30530678

    Open questions at the time
    • Switch between K48 and K63 output not mechanistically explained
    • Recruitment to damaged mitochondria incompletely defined
  12. 2018 Medium

    Dissected domain contributions, showing the RING domain mediates Ccr4-Not/Pol II association while the RRM-C domain controls a distinct gene subset, with proteasomal defects requiring both.

    Evidence Domain mutagenesis, RNA-seq, Pol II ChIP and proteasome assays in yeast

    PMID:29802328

    Open questions at the time
    • RRM-C RNA targets not identified
    • Integration of catalytic and RNA-binding functions unclear
  13. 2019 High

    Established the metazoan binding mode to CCR4-NOT through a conserved CAF40-binding motif, structurally defining a mutually exclusive interface and linking it to functional mRNA decay.

    Evidence Crystal structures, binding assays, mutagenesis and tethered reporter decay

    PMID:30692204

    Open questions at the time
    • Endogenous mRNA targets recruited via this motif unknown
    • Relationship to ligase activity not addressed
  14. 2021 High

    Connected CNOT4 to translation elongation, mRNA adaptor function, ubiquitin-proteasome degradation steps, and stress-induced cytoplasmic localization, broadening its physiological reach to meiosis and codon-dependent elongation.

    Evidence Ribosome profiling, conditional KO mouse, UFD substrate and ubiquitin-conjugate binding assays, and localization analysis

    PMID:33727038 PMID:34026442 PMID:34338747 PMID:34469733

    Open questions at the time
    • Mechanism coupling elongation control to ligase activity incomplete
    • How cytoplasmic foci relate to function undefined
  15. 2023 Medium

    Linked Not4-driven ribosome pausing to co-translational mitochondrial mRNA docking and showed Not1 vs Not4 inversely control mRNA solubility tied to codon optimality, refining its role in spatial mRNA-protein homeostasis.

    Evidence Ribosome profiling of soluble/insoluble fractions, mitochondrial fractionation and genetic epistasis in yeast

    PMID:36803582 PMID:37094076

    Open questions at the time
    • Direct substrate(s) driving pausing not fully defined
    • Generality beyond MMF1 unclear
  16. 2023 Medium

    Extended degradative substrates to disease-relevant human targets PAF1 and YAP in cancer cells, including a small-molecule (hydnocarpin) exploiting the CNOT4-YAP axis.

    Evidence Co-IP, ubiquitination assays, MST/DARTS binding in NSCLC and TNBC cells

    PMID:37493105 PMID:37802875

    Open questions at the time
    • YAP degradation mechanistic detail limited (Low-confidence for the YAP/drug finding)
    • In vivo relevance of YAP axis not established
  17. 2024 Medium

    Identified ZNF598 as a CNOT4 substrate during ribosome quality control and showed CNOT4 itself undergoes regulatory K63 and TRIM21/TNKS1BP1-mediated K48/K6 ubiquitination, embedding it in feedback regulation.

    Evidence Linkage-specific in vivo ubiquitination, site-specific mutants, Drosophila genetics and Co-IP

    PMID:38388640 PMID:39019859

    Open questions at the time
    • Cross-talk between CNOT4 auto-ubiquitination and substrate selection unknown
    • Stoichiometry and turnover kinetics undefined
  18. 2025 Medium

    Resolved the human CCR4-NOT relationship as non-constitutive and showed CNOT4 depletion accelerates mRNA decay opposite to CNOT1, while expanding regulatory roles to PPARγ-dependent adipogenesis, crinophagy, and TENT5C destabilization.

    Evidence Auxin-degron depletion with BioID and transcriptome decay, ChIP in Cnot4-Het mice, Drosophila imaging and proteomic stability assays (two preprints)

    PMID:40424271 PMID:41161383 PMID:41857610 PMID:bio_10.1101_2024.11.14.623596 PMID:bio_10.1101_2025.09.24.678350

    Open questions at the time
    • Mechanism of the opposite mRNA-decay effect versus CNOT1 unexplained
    • TENT5C and crinophagy findings are preprints with limited CNOT4 mechanistic detail

Open questions

Synthesis pass · forward-looking unresolved questions
  • How CNOT4 selects among its many substrates and switches between K48, K63, and monoubiquitination outputs in different cellular contexts remains unresolved.
  • No unifying model for substrate/linkage specificity
  • Relationship between ligase activity and CCR4-NOT-associated mRNA functions unclear
  • Human-cell substrate repertoire incompletely mapped

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0016874 ligase activity 7 GO:0140096 catalytic activity, acting on a protein 7 GO:0003723 RNA binding 2 GO:0060090 molecular adaptor activity 2 GO:0140110 transcription regulator activity 2
Localization
GO:0005840 ribosome 3 GO:0005634 nucleus 2 GO:0005829 cytosol 2
Pathway
R-HSA-392499 Metabolism of proteins 5 R-HSA-8953854 Metabolism of RNA 4 R-HSA-74160 Gene expression (Transcription) 3 R-HSA-8953897 Cellular responses to stimuli 3 R-HSA-9612973 Autophagy 2
Partners
CAF40/CNOT9CNOT1CNOT7ECM29TNKS1BP1TRIM21UBCH5BUSP4
Complex memberships
CCR4-NOT complex

Evidence

Reading pass · 33 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1994 NOT4 physically interacts with NOT1 and NOT3 in the yeast two-hybrid assay, and NOT1/NOT2/NOT3/NOT4 form a ~500 kDa nuclear complex that acts as a global negative regulator of RNA Pol II transcription, preferentially repressing TC-element-dependent transcription. Two-hybrid interaction, biochemical co-fractionation, allele-specific suppression genetics Genes & development Medium 7926748
1999 In the yeast CCR4-NOT complex, NOT4 (and NOT3) interact with the C-terminal region (residues 1490–2108) of NOT1, placing NOT4 peripheral to the CCR4-CAF1-NOT1-(NOT2,NOT5) core; NOT4 and NOT3 do not significantly affect the association of CCR4, CAF1, and NOT1 with each other. Co-immunoprecipitation, two-hybrid, deletion mapping, genetic analysis Molecular and cellular biology Medium 10490603
2000 Human NOT4 contains a C4C4-type RING finger (residues 1–78) that coordinates two zinc ions in a cross-brace manner via cysteine residues; the NMR solution structure shows an alpha-helix and three long loops, overall similar to C3HC4 RING fingers but with distinct secondary structural elements in non-conserved regions. Heteronuclear NMR structure determination with 113Cd-substitution to confirm cross-brace zinc coordination The Journal of biological chemistry High 11087754
2004 The N-terminal RING finger domain of CNOT4 acts as an E3 ubiquitin ligase that specifically interacts with the ubiquitin-conjugating enzyme UbcH5B; NMR chemical shift perturbation mapping identified UbcH5B residues contacting the CNOT4 RING domain, and HADDOCK docking revealed the structural basis of E2/E3 specificity differing from the c-Cbl/UbcH7 interface. NMR chemical shift perturbation, HADDOCK computational docking, biochemical mutagenesis Structure High 15062086
2005 The two zinc-binding sites within the C4C4 RING domain of CNOT4 show differential metal exchange rates; NMR backbone dynamics analysis shows this differential stability arises from differences in site accessibility and electrostatic interactions, and CNOT4 RING is less flexible than the p44 RING domain. NMR backbone dynamics (Zn2+/Cd2+ exchange), relaxation measurements Journal of molecular biology Medium 15890366
2010 Not4, together with Ubc4, is required for ubiquitin-mediated destabilization of Cdc17 (the catalytic subunit of DNA polymerase-α) in yeast; disruption of this turnover pathway causes hydroxyurea sensitivity and synthetic dosage lethality upon Cdc17 overexpression. Genetic deletion analysis, protein stability assays, growth phenotype assays (HU sensitivity, SDL) Molecular biology of the cell Medium 20660159
2011 Not4 E3 ligase is required for proper proteasome assembly: in not4Δ cells the regulatory particle (RP) and core particle (CP) form aberrant salt-resistant complexes, and Ecm29 (a proteasome chaperone) becomes ubiquitinated and degraded. Not4 associates with RP species in purified proteasome holoenzyme and interacts with Ecm29 to maintain normal proteasome integrity. Co-immunoprecipitation, proteasome purification/fractionation, in vivo ubiquitination assay, deletion genetics Molecular and cellular biology High 21321079
2012 Not4 ubiquitin ligase ubiquitinates Rps7A (but not its paralogue Rps7B) in vivo and in vitro; ubiquitinated Rps7A is found only in 80S and polysome fractions. Not4 is itself detectable in polysome fractions, and optimal presence of Not5 in polysomes depends on Not4. In vitro ubiquitination assay, polysome fractionation/sedimentation, deletion genetics, protein aggregation analysis Molecular microbiology High 22243599
2012 Not4p ubiquitin ligase mediates H2O2-induced destruction of cyclin C via K48-linked polyubiquitination, directing it to the 26S proteasome. Cyclin C must translocate from nucleus to cytoplasm (requiring the cell-wall-integrity MAPK module and phospholipase C) before Not4p-dependent degradation; a cytoplasm-restricted cyclin C derivative is still subject to Not4p-dependent destruction. In vitro polyubiquitination assay with K48 linkage-specific analysis, genetic deletion, subcellular localization by microscopy, epistasis with MAPK/PLC pathway mutants Journal of cell science High 22421358
2012 The nuclear E3 ubiquitin ligase Not4 binds to Yap1 in an oxidant-stimulated manner and is required for proteasome-dependent Yap1 degradation after oxidative stress; Yap1 degradation requires nuclear localization and DNA-binding ability of Yap1 but not chromatin binding. Co-immunoprecipitation, genetic screen of ubiquitin pathway mutants, proteasome inhibitor treatment, localization-restricted Yap1 mutant analysis The Journal of biological chemistry Medium 22707721
2014 Not4 associates with polysomes and contributes to translational repression of transcripts that cause transient ribosome stalling; absence of Not4 impairs global translational repression upon nutrient withdrawal, enhances expression of arrested nascent polypeptides, and causes constitutive protein folding stress and aggregation. Polysome fractionation, protein aggregation assays, pulse-chase translation analysis, genetic epistasis with decapping machinery mutants The EMBO journal Medium 25971775
2014 Not4 promotes protein quality control independently of the Ccr4 deadenylase module; clearance of aberrant proteins by Not4 operates at least in part via the proteasome. Genetic deletion of NOT4 vs CCR4, protein aggregation assays, proteasome activity measurements, epistasis analysis PloS one Medium 24465968
2015 Human CNOT4 controls PAF1 protein levels by promoting K48-linked polyubiquitination of PAF1 and its subsequent degradation by the 26S proteasome; this degradation requires nuclear localization of PAF1 but not chromatin binding. Co-immunoprecipitation, ubiquitination assay, proteasome inhibitor treatment, NLS-deficient PAF1 mutant analysis PloS one Medium 25933433
2017 CNOT4 acts as an E3 ubiquitin ligase for influenza A virus nucleoprotein (NP): CNOT4 interacts with NP in cells and ubiquitinates NP in vitro without affecting NP protein stability; monoubiquitination at residues K184, K227, and K273 in the RNA-binding groove of NP promotes viral RNA replication. Co-immunoprecipitation, in vitro ubiquitination assay, CNOT4 knockdown/overexpression, mass spectrometry ubiquitination site mapping, arginine substitution mutagenesis mBio High 28536288
2018 Not4 selectively ubiquitinates Rpt5 during proteasomal ATPase (Rpt ring) assembly; Not4 competes with chaperones Nas2 and Hsm3 for access to Rpt5 ubiquitination sites, which are sterically occluded by these chaperones. Rpt5 ubiquitination does not destabilize the ring but blocks incorporation of Rpn1 ubiquitin receptor and Ubp6 deubiquitinase, thereby acting as an assembly checkpoint. In vitro ubiquitination reconstitution, structural modeling using known Rpt-chaperone co-crystal structures, proteasome assembly intermediate analysis, deletion/competition biochemistry Proceedings of the National Academy of Sciences of the United States of America High 30530678
2018 Upon mitochondrial damage, CNOT4 is recruited to the stalled ribosome/mRNA complex on the mitochondrial outer membrane and ubiquitinates ABCE1; this K63-linked poly-ubiquitin signal on ABCE1 recruits autophagy receptors to the mitochondrial outer membrane to initiate mitophagy downstream of PINK1 signaling. Co-immunoprecipitation, in vivo ubiquitination assay, genetic epistasis in Drosophila PINK1 model, ribosome fractionation Cell metabolism High 29861391
2018 The Not4 RING domain is required for Ccr4-Not association with RNA Pol II, while the conserved RRM-C domain (RNA recognition motif and C3H1 domain) affects a specific subset of Pol II-regulated genes without affecting Ccr4-Not/Pol II association; individual RING or RRM-C mutations fail to replicate proteasomal defects seen in not4Δ cells, but combined RING/RRM-C mutations do. Domain mutagenesis, transcriptome analysis (RNA-seq), ChIP (Pol II occupancy), proteasome integrity assay, Co-IP Scientific reports Medium 29802328
2019 Metazoan NOT4 contains a conserved CAF40-binding motif (CBM) in its C-terminal region that directly binds the CAF40 subunit of the CCR4-NOT complex; crystal structures of the CBM-CAF40 complex show this binding surface is mutually exclusive with Roquin or Bag of marbles binding. CAF40 depletion or structure-guided mutagenesis of the NOT4-CAF40 interface impairs NOT4-dependent decay of tethered reporter mRNAs. Crystal structure determination, NMR/binding assays, mutagenesis, tethered mRNA reporter decay assay, CAF40 depletion Genes & development High 30692204
2021 Not4 and Not5 modulate translation elongation dynamics and change ribosome A-site dwelling occupancy in a codon-dependent fashion; Not4 E3 ligase activity (Rps7A ubiquitination) and a moonlighting function of Rli1 enable Not4-dependent translation of polyarginine stretches. Not5 resides in punctate loci (condensates) that co-purify with ribosomes and Rli1 but exclude eIF5A. Ribosome profiling, polysome fractionation, in vivo ubiquitination, microscopy, genetic deletion and epistasis Cell reports High 34469733
2021 In male germ cells, CNOT4 functions as an mRNA adaptor of the CCR4-NOT complex by targeting mRNAs to CNOT7 for deadenylation/degradation; conditional Cnot4 knockout in mouse male germ cells causes defective DNA double-strand break repair, impaired XY chromosome crossover, and infertility. Conditional knockout mouse model, RNA-seq, poly(A) tail analysis, immunofluorescence for meiotic markers Advanced science High 34026442
2021 Ccr4, Caf1, and Not4 function at a post-ubiquitylation step in the ubiquitin-proteasome system to promote degradation of ubiquitinated substrates; Not4, Ccr4, and Caf1 bind cellular ubiquitin conjugates, and loss of any of these three genes impairs UPS-mediated degradation of a UFD pathway substrate in yeast. UFD pathway substrate degradation assays, ubiquitin conjugate binding assays, genetic deletion, proteasome interaction assays Biochimica et biophysica acta. Molecular cell research Medium 33727038
2021 Yeast Not4 localizes to cytoplasmic foci after cellular stress; the C-terminal 211 amino acids are required for proper localization to stress-induced cytoplasmic foci and for Not4 function in stress response gene regulation, while the last 120 amino acids are dispensable. Fluorescence microscopy (subcellular localization), domain deletion analysis, stress response gene expression assays FEMS microbiology letters Medium 34338747
2023 Not4 promotes ribosome pausing during MMF1 mRNA translation, enabling co-translational docking of MMF1 mRNA to mitochondria via the MMF1 nascent chain mitochondrial targeting sequence, Egd1 chaperone, Om14, and the co-translational import machinery; Not4 also ubiquitinates the Egd1 chaperone, and this pathway (Mito-ENCay) additionally requires Caf130, Cis1, autophagy, and no-go-decay to limit MMF1 overexpression. Ribosome profiling, polysome fractionation, in vivo ubiquitination assay, mitochondrial fractionation, genetic epistasis with autophagy/no-go-decay mutants Nucleic acids research Medium 37094076
2023 Depletion of Not1 and Not4 inversely affects mRNA solubility: depletion of Not4 solubilizes mRNAs with lower non-optimal codon content and higher expression, while depletion of Not1 solubilizes mitochondrial mRNAs; insoluble mRNAs are enriched for ribosomes dwelling at non-optimal codons and show higher proportion of co-translational degradation. Ribosome profiling of soluble vs insoluble mRNA fractions, transcriptome-wide mRNA decay analysis, genetic depletion Genome biology Medium 36803582
2023 CNOT4 facilitates ubiquitination and proteasomal degradation of PAF1 in non-small cell lung cancer cells, as confirmed by co-immunoprecipitation of CNOT4 with PAF1 and in vivo ubiquitination assays. Co-immunoprecipitation, ubiquitination assay, proteasome inhibitor treatment, CNOT4 overexpression in NSCLC cells Molecular carcinogenesis Medium 37493105
2023 CNOT4 directly binds YAP and promotes its ubiquitination and proteasomal degradation; the drug hydnocarpin binds both YAP and CNOT4 (confirmed by microscale thermophoresis and DARTS assay) and exploits this pathway to degrade YAP in triple negative breast cancer cells. Microscale thermophoresis, DARTS assay, ubiquitination assay, proteasome inhibitor rescue, co-immunoprecipitation China journal of Chinese materia medica Low 37802875
2024 CNOT4 undergoes regulatory K63-linked ubiquitination in a CNOT4-dependent manner (auto-ubiquitination context) and ubiquitinates ZNF598; this ubiquitination is required for ZNF598-mediated resolution of stalled ribosomes and protection against mitochondrial stress. In vivo ubiquitination assay with K63-linkage specificity, CNOT4 overexpression/depletion, Drosophila genetic models, ribosome stalling assays Nature communications Medium 38388640
2024 TNKS1BP1 interacts with TRIM21 and mediates ubiquitination of CNOT4 at K239 via K48 and K6 linkage, leading to CNOT4 degradation; this promotes inhibition of JAK2/STAT3 pathway activity, thereby inducing lipid accumulation and inhibiting autophagy in hepatocellular carcinoma cells. Co-immunoprecipitation, ubiquitination assay with site-specific mutant (K239), western blotting, genetic knockdown Cell death & disease Medium 39019859
2025 Human CNOT4 does not form a stable complex with CCR4-NOT by standard biochemical co-purification but associates with the complex in cells as shown by BioID proximity labeling; depletion of CNOT4 does not affect CCR4-NOT complex integrity but paradoxically accelerates global mRNA decay, opposite to the effect of CNOT1 depletion. mRNA stability changes in CCR4-NOT-depleted cells correlate with codon optimality. Auxin-induced degron depletion, BioID proximity labeling, transcriptome-wide mRNA abundance and decay measurement, co-immunoprecipitation The Journal of biological chemistry Medium 41161383
2025 In Drosophila salivary gland cells, Cnot4 ubiquitin ligase mediates ubiquitination of secretory granules (glue granules), marking them for crinophagic degradation (lysosomal fusion); loss of Cnot4 impairs glue granule fusion with lysosomes, while Cnot4 overexpression induces premature crinophagy. Genetic loss-of-function, overexpression, fluorescence microscopy of granule-lysosome fusion, in vivo ubiquitination bioRxivpreprint Medium bio_10.1101_2025.09.24.678350
2025 In mouse/human cells, CNOT4 ubiquitinates and destabilizes TENT5C poly(A) polymerase, providing a mechanism by which the CCR4-NOT deadenylase complex limits the stability of a cytoplasmic polyadenylation enzyme during erythropoiesis. Proteomic interaction assay, ubiquitination/protein stability analysis, genetic depletion bioRxivpreprint Low bio_10.1101_2024.11.14.623596
2025 USP4 deubiquitinase directly binds PAF1, removes K48-linked polyubiquitin chains added by CNOT4, and thereby stabilizes PAF1 protein levels; this antagonism between USP4 and CNOT4 controls PAF1 abundance and NSCLC tumor growth in vivo. Co-immunoprecipitation, K48-linkage-specific ubiquitination assay, proteasome inhibition, xenograft mouse model Respiratory research Medium 41857610
2025 Heterozygous deletion of Cnot4 in mice impairs PPARγ transcriptional activity: Cnot4 Het fibroblasts show decreased PPARγ binding to target gene promoters in response to rosiglitazone, reducing adipogenic gene expression and adipocyte differentiation capacity. Cnot4 heterozygous knockout mouse, ChIP for PPARγ promoter binding, gene expression analysis, in vitro adipocyte differentiation assay PloS one Medium 40424271

Source papers

Stage 0 corpus · 39 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1994 NOT1(CDC39), NOT2(CDC36), NOT3, and NOT4 encode a global-negative regulator of transcription that differentially affects TATA-element utilization. Genes & development 183 7926748
1999 The CCR4 and CAF1 proteins of the CCR4-NOT complex are physically and functionally separated from NOT2, NOT4, and NOT5. Molecular and cellular biology 137 10490603
2004 Structural model of the UbcH5B/CNOT4 complex revealed by combining NMR, mutagenesis, and docking approaches. Structure (London, England : 1993) 111 15062086
2012 Presence of Not5 and ubiquitinated Rps7A in polysome fractions depends upon the Not4 E3 ligase. Molecular microbiology 79 22243599
2018 Ubiquitination of ABCE1 by NOT4 in Response to Mitochondrial Damage Links Co-translational Quality Control to PINK1-Directed Mitophagy. Cell metabolism 78 29861391
2000 The structure of the C4C4 ring finger of human NOT4 reveals features distinct from those of C3HC4 RING fingers. The Journal of biological chemistry 77 11087754
2011 Not4 E3 ligase contributes to proteasome assembly and functional integrity in part through Ecm29. Molecular and cellular biology 68 21321079
2012 Oxidative-stress-induced nuclear to cytoplasmic relocalization is required for Not4-dependent cyclin C destruction. Journal of cell science 53 22421358
2017 CNOT4-Mediated Ubiquitination of Influenza A Virus Nucleoprotein Promotes Viral RNA Replication. mBio 47 28536288
2015 Not4-dependent translational repression is important for cellular protein homeostasis in yeast. The EMBO journal 46 25971775
2012 Proteolytic degradation of the Yap1 transcription factor is regulated by subcellular localization and the E3 ubiquitin ligase Not4. The Journal of biological chemistry 42 22707721
2014 The Not4 E3 ligase and CCR4 deadenylase play distinct roles in protein quality control. PloS one 39 24465968
2019 A conserved CAF40-binding motif in metazoan NOT4 mediates association with the CCR4-NOT complex. Genes & development 35 30692204
2021 Not4 and Not5 modulate translation elongation by Rps7A ubiquitination, Rli1 moonlighting, and condensates that exclude eIF5A. Cell reports 33 34469733
2021 The CNOT4 Subunit of the CCR4-NOT Complex is Involved in mRNA Degradation, Efficient DNA Damage Repair, and XY Chromosome Crossover during Male Germ Cell Meiosis. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 31 34026442
2004 Deletion of the NOT4 gene impairs hyphal development and pathogenicity in Candida albicans. Microbiology (Reading, England) 31 14702416
2014 The role of the E3 ligase Not4 in cotranslational quality control. Frontiers in genetics 29 24904641
2010 Ubc4 and Not4 regulate steady-state levels of DNA polymerase-α to promote efficient and accurate DNA replication. Molecular biology of the cell 23 20660159
2005 Dynamics and metal exchange properties of C4C4 RING domains from CNOT4 and the p44 subunit of TFIIH. Journal of molecular biology 22 15890366
2013 The Not4 RING E3 Ligase: A Relevant Player in Cotranslational Quality Control. ISRN molecular biology 16 27335678
2024 Stalled translation by mitochondrial stress upregulates a CNOT4-ZNF598 ribosomal quality control pathway important for tissue homeostasis. Nature communications 15 38388640
2015 Protein Degradation of RNA Polymerase II-Association Factor 1(PAF1) Is Controlled by CNOT4 and 26S Proteasome. PloS one 14 25933433
2018 Ubiquitin-dependent switch during assembly of the proteasomal ATPases mediated by Not4 ubiquitin ligase. Proceedings of the National Academy of Sciences of the United States of America 13 30530678
2024 TNKS1BP1 facilitates ubiquitination of CNOT4 by TRIM21 to promote hepatocellular carcinoma progression and immune evasion. Cell death & disease 12 39019859
2023 Not1 and Not4 inversely determine mRNA solubility that sets the dynamics of co-translational events. Genome biology 12 36803582
2018 The conserved RNA recognition motif and C3H1 domain of the Not4 ubiquitin ligase regulate in vivo ligase function. Scientific reports 11 29802328
2021 Aberrant expression of MYD88 via RNA-controlling CNOT4 and EXOSC3 in colonic mucosa impacts generation of colonic cancer. Cancer science 10 34626022
2023 CNOT4 suppresses nonsmall cell lung cancer progression by promoting the degradation of PAF1. Molecular carcinogenesis 6 37493105
2020 CNOT4 enhances the efficacy of anti-PD-1 immunotherapy in a model of non-small cell lung cancer. FEBS open bio 6 33034149
2023 Not4-dependent targeting of MMF1 mRNA to mitochondria limits its expression via ribosome pausing, Egd1 ubiquitination, Caf130, no-go-decay and autophagy. Nucleic acids research 4 37094076
2023 [Hydnocarpin inhibits malignant progression of triple negative breast cancer via CNOT4-mediated ubiquitination and degradation of YAP]. Zhongguo Zhong yao za zhi = Zhongguo zhongyao zazhi = China journal of Chinese materia medica 4 37802875
2021 CNOT4 suppresses non-small cell lung cancer progression and is required for effector cytolytic T lymphocytes cell responses to lung cancer cells. Molecular immunology 4 33592572
2021 Ccr4-Not complex subunits Ccr4, Caf1, and Not4 are novel proteolysis factors promoting the degradation of ubiquitin-dependent substrates by the 26S proteasome. Biochimica et biophysica acta. Molecular cell research 4 33727038
2025 Cnot4 heterozygosity attenuates high fat diet-induced obesity in mice and impairs PPARγ-mediated adipocyte differentiation. PloS one 3 40424271
2025 Auxin-induced depletion of human CCR4-NOT subunits reveals opposing functions of CNOT1 and CNOT4 in mRNA metabolism. The Journal of biological chemistry 2 41161383
2023 Depletion of CNOT4 modulates the DNA damage responses following ionizing radiation (IR). Journal of cancer research and therapeutics 1 38554309
2026 USP4 promotes non-small cell lung cancer tumorigenesis by antagonizing CNOT4-mediated degradation of PAF1. Respiratory research 0 41857610
2024 Ubiquitination-deficit of Cnot4 impairs the capacity of proliferation and differentiation in mouse embryonic stem cells. Biochemical and biophysical research communications 0 39798536
2021 The C-terminal region of yeast ubiquitin-protein ligase Not4 mediates its cellular localization and stress response. FEMS microbiology letters 0 34338747

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