| 2023 |
Cryo-EM structure of the RNF20/RNF40-hRAD6A-Ub-nucleosome complex (captured via chemical trapping) revealed that RNF40 directly binds nucleosomal DNA and exhibits a conserved E3/E2/nucleosome interaction pattern for H2B monoubiquitylation at K120; an uncanonical non-hydrophobic contact in the RING-E2 interface positions RAD6A directly above the target H2B lysine residue. |
Chemical trapping strategy + cryo-EM structure determination + mutagenesis |
Molecular cell |
High |
37633270
|
| 2016 |
Crystal structure of the RNF20 RING domain shows it forms a homodimer that specifically interacts with the Ube2B~Ub conjugate; the RING domains of RNF20 and RNF40 can form a stable heterodimer that is catalytically active; key contacts at the E3-E2 and E3-ubiquitin interfaces were identified by mutagenesis. |
X-ray crystallography + mutagenesis of E3-E2 and E3-ubiquitin interfaces + in vitro ubiquitination assay |
Journal of molecular biology |
High |
27569044
|
| 2026 |
Cryo-EM structure of RNF20/RNF40-RAD6A-Ub-H2BS112GlcNAc nucleosome complex showed that H2B S112 GlcNAcylation interacts with the E2 enzyme RAD6A (not the E3 RNF20/RNF40), allosterically enhancing the nucleophilicity of H2B K120 to stimulate ubiquitin transfer; structure-activity relationship identified the C2 N-acetyl group and β-configuration of C1 as essential. |
Chemical synthesis of modified nucleosomes + cryo-EM + mutagenesis + kinetics assays |
Nature chemical biology |
High |
41495224
|
| 2026 |
Crystal structure of the Bre1-Lge1 complex and AlphaFold model of RNF20/RNF40-WAC revealed extensive interaction interfaces; in vitro and in vivo experiments demonstrated that RNF20/RNF40-WAC interactions are critical for H2B monoubiquitination, with different electrostatic contacts encoding binding specificity compared to yeast Bre1-Lge1. |
X-ray crystallography (Bre1-Lge1) + AlphaFold modeling (RNF20/RNF40-WAC) + in vitro ubiquitination assay + in vivo functional experiments |
Nucleic acids research |
High |
41533567
|
| 2012 |
RNF20/RNF40 (mammalian Bre1 complex) functions as the E3 ubiquitin ligase for histone H2B monoubiquitination; deficiency leads to aberrant R-loop formation causing replication-associated double-strand breaks, combined with a defect in homologous recombination, resulting in chromosomal instability. |
Loss-of-function (Bre1-deficient cells) + detection of R-loops, DSBs, and genomic rearrangements |
Cancer research |
High |
22354749
|
| 2011 |
RNF20 and RNF40 form a heterodimeric E3 ubiquitin ligase complex that monoubiquitinates histone H2B at lysine 120; the tumor suppressor CDC73 interacts with both RNF20 and RNF40 at discrete but closely located residues and is required for maintenance of H2Bub1 levels. |
Yeast two-hybrid + co-immunoprecipitation + knockdown of CDC73 + western blot for H2Bub1 |
Human molecular genetics |
High |
22021426
|
| 2011 |
RNF40 depletion results in sustained γH2AX phosphorylation, decreased rapid cell cycle checkpoint activation, decreased H3K56ac, and decreased recruitment of the FACT complex (specifically SUPT16H) to chromatin following DNA DSBs; RNF40 and SUPT16H cooperate to promote DNA end resection and timely DNA repair. |
siRNA knockdown + immunofluorescence for γH2AX + western blot for H3K56ac + chromatin fractionation + DNA end resection assay |
Cell cycle |
High |
22031019
|
| 2014 |
Arsenite directly binds to the RING finger domains of RNF20 and RNF40 via cysteine residues in vitro and in cells, inhibiting H2B ubiquitination and impairing recruitment of BRCA1 and RAD51 to DNA DSB sites, thereby compromising DNA DSB repair. |
In vitro binding assay + cellular arsenite treatment + ChIP for BRCA1/RAD51 recruitment + comet assay/western blot for DSB repair |
Journal of the American Chemical Society |
High |
25170678
|
| 2019 |
RNF20 and RNF40 are required for DSB repair via both homologous recombination and class switch recombination (NHEJ) in mouse B cells; DSBs induce a global increase in H2Bub1 regulated jointly by ATM and ATR kinases, independently of H2AX phosphorylation. |
RNF20/RNF40 knockout in mouse B cells + class switch recombination assay + HR assay + western blot + ATM/ATR inhibitor treatment |
Molecular and cellular biology |
High |
30692271
|
| 2011 |
RNF20 and RNF40 physically and functionally interact with the androgen receptor (AR) and modulate its transcriptional activity; androgen induction of FKBP51 and PSA is accompanied by dynamic increases in H2Bub1 within transcribed regions of these loci. |
Co-immunoprecipitation + reporter assay + ChIP for H2Bub1 at AR target loci |
Molecular and cellular endocrinology |
Medium |
22155569
|
| 2017 |
RNF20, RNF40, and WAC form an E3 ligase complex that mediates H2B ubiquitination; knockdown of WAC phenocopies loss of H2B ubiquitination and causes cell death in MLL-rearranged ALL cells. |
siRNA knockdown + western blot for H2Bub1 + cell viability assay |
Leukemia |
Medium |
28690313
|
| 2019 |
RNF40 loss decreases nuclear localization of NF-κB following TNF-α treatment in colorectal cancer cells, reducing induction of NF-κB-associated cytokines; colon-specific Rnf40 deletion exerts a protective effect in a murine model of acute colitis. |
siRNA knockdown + immunofluorescence for NF-κB nuclear localization + mRNA-seq + in vivo colitis model with colon-specific Rnf40 knockout |
Journal of Crohn's & colitis |
Medium |
30321325
|
| 2019 |
RNF40 depletion in colorectal cancer cells increases apoptosis via elevated caspase 3/7 activity, associated with reduced anti-apoptotic and elevated pro-apoptotic BCL2 family member expression; H2Bub1 directly occupies transcribed regions of anti-apoptotic genes. |
siRNA knockdown + flow cytometry (cell cycle) + caspase 3/7 assay + Annexin V + ChIP-seq for H2Bub1 + mRNA-seq + caspase inhibitor rescue |
Clinical epigenetics |
Medium |
31266541
|
| 2020 |
RNF40-driven H2B monoubiquitination is essential for transcriptional activation of RHO/ROCK/LIMK pathway components and proper actin-cytoskeleton dynamics through trans-histone crosstalk with H3K4me3 in HER2+ breast cancer. |
Tissue-specific Rnf40 deletion (mouse mammary carcinoma model) + ChIP for H2Bub1 and H3K4me3 + transcriptome analysis + actin dynamics assays |
Cell death & disease |
Medium |
33070155
|
| 2020 |
RNF40 loss impairs early gene activation during somatic cell reprogramming to iPSCs; mechanistically, RNF40 controls expression of the PRC2 component EZH2 and promotes resolution of H3K4me3/H3K27me3 bivalency on H2Bub1-occupied pluripotency genes. |
RNF40 knockdown + iPSC reprogramming assay + ChIP-seq for H3K4me3, H3K27me3, H2Bub1 + mRNA-seq |
Cell death & disease |
Medium |
32341358
|
| 2020 |
Osteoblast-specific RNF40 deletion in mice impairs early lineage specification, decreases osteoclast number and function via reduced RANKL (Tnfsf11) expression in osteoblasts; H2B monoubiquitination directly occupies the Tnfsf11 locus. |
Conditional osteoblast-specific Rnf40 knockout mouse + bone phenotype analysis + ChIP for H2Bub1 at Tnfsf11 + RANKL expression analysis |
Cell death and differentiation |
Medium |
32901120
|
| 2021 |
Intestine-specific deletion of Rnf20 or Rnf40 in mice causes spontaneous colorectal inflammation; mechanistically, RNF20/RNF40-mediated H2Bub1 controls H3K4me3 occupancy and transcription of the Vitamin D Receptor (Vdr) gene and its target genes in intestinal epithelial cells. |
Conditional intestinal Rnf20/Rnf40 knockout + mRNA-seq + ChIP-seq for H2Bub1 and H3K4me3 + in vivo colitis monitoring |
Cell death and differentiation |
Medium |
34088983
|
| 2024 |
RNF40 mediates ubiquitination and proteasome-dependent degradation of LIMA1 (an actin-binding protein involved in cholesterol absorption) as a non-histone substrate; the 1-166aa fragment of LIMA1 is required for interaction with RNF40, and RNF40 overexpression reduces cellular lipid content in a LIMA1-dependent manner. |
Co-immunoprecipitation + domain mapping + ubiquitination assay + proteasome inhibitor treatment + lipid content measurement + rescue by LIMA1 overexpression |
Cell death discovery |
Medium |
38909032
|
| 2025 |
RNF40 catalyzes K6- and K11-linked polyubiquitination of KDM6A, targeting it for autophagic degradation via TAX1BP1 recognition; this contrasts with USP7-mediated deubiquitination (K48-linked) that prevents KDM6A proteasomal degradation, forming an antagonistic ubiquitin-switching circuit regulating KDM6A stability and coronavirus receptor expression. |
Genetic and pharmacological inhibition + ubiquitination linkage analysis + co-immunoprecipitation + autophagic degradation assay + viral infection functional assay |
Advanced science |
Medium |
41691482
|
| 2025 |
RNF40 promotes K48-linked polyubiquitination and proteasomal degradation of Parkin in cerebrovascular endothelial cells, thereby inhibiting mitophagy; Rnf40 knockdown in spontaneously hypertensive rats restored mitophagy, improved tight junction proteins, and protected the blood-brain barrier. |
In vivo Rnf40 knockdown (pAAV-shRnf40 in SHRs) + ubiquitination assay (K48 linkage) + mitophagy assay + tight junction protein western blot + cerebral blood flow measurement |
CNS neuroscience & therapeutics |
Medium |
39777866
|
| 2023 |
The RNF40/H2Bub1 axis positively regulates peroxisome-related gene expression (PRDX5, PEX6, PMVK); loss of RNF20/RNF40 impairs peroxisomal biogenesis and ROS metabolism, leading to increased lipid peroxidation and ferroptotic cell death in cervical cancer cells. |
RNF20/RNF40 knockdown + ChIP-qPCR for H2Bub1 + transcriptome analysis + lipid peroxidation assay + flow cytometry for ferroptosis markers + in vivo CAM assay |
Cell communication and signaling |
Medium |
40597205
|
| 2023 |
The RNF40/H2Bub1 axis promotes cancer stem cell properties and glycolytic program in triple-negative breast cancer by supporting YAP1 signaling. |
RNF40 knockdown/overexpression + sphere formation assay + ChIP-seq for H2Bub1 + transcriptome analysis + YAP1 pathway analysis |
Cell death & disease |
Medium |
37770435
|
| 2025 |
PAF1C-driven transcription restart after DNA damage is independent of RNF20/RNF40-mediated H2B-K120 ubiquitination; H2Bub1 levels do not correlate with transcription restoration following DNA lesions, and RTF1-stimulated H2Bub1 does not contribute to restart. |
RNF20/RNF40 depletion + transcription restart assay after DNA damage + H2Bub1 ChIP |
bioRxivpreprint |
Low |
bio_10.1101_2025.07.23.666359
|
| 2024 |
RNF40 physically interacts with RTF1 and is required for histone H2B monoubiquitination that supports Th17 cell differentiation; RNF40-deficient cells phenocopy RTF1-deficient cells in impaired Th17 differentiation. |
RNF40 knockout in T cells + Th17 differentiation assay + H2Bub1 ChIP-seq + genetic epistasis with RTF1 |
bioRxivpreprint |
Low |
bio_10.1101_2024.11.08.622668
|