| 2011 |
Ube2w acts as an initiator E2 that monoubiquitinates CHIP (the E3 ligase), and this monoubiquitination stabilizes the interaction between CHIP and the deubiquitinase ataxin-3; ataxin-3 then limits polyubiquitin chain length on CHIP substrates and subsequently deubiquitinates CHIP to terminate the ubiquitination cycle. |
In vitro ubiquitination assays, co-immunoprecipitation, and functional DUB activity assays |
Molecular cell |
High |
21855799
|
| 2013 |
Ube2W (Ubc16) is an E2 ubiquitin-conjugating enzyme with specific protein N-terminal mono-ubiquitylation activity; it conjugates ubiquitin to the α-amino group of protein N-termini (including its own N-terminus and that of SUMO) rather than to lysine ε-amino groups. |
In vitro ubiquitylation assays, mass spectrometry identification of modification site, and biochemical assays with RNF4 |
The Biochemical journal |
High |
23560854 23696636
|
| 2013 |
Ube2w ubiquitinates the N-terminal α-NH2 group of substrates (not internal lysines); this was demonstrated using lysine-less and N-terminally blocked versions of ataxin-3 and Tau, and confirmed by direct N-terminal sequencing of ubiquitinated ataxin-3; Ube2w contains novel active-site residues important for this activity. |
Mass spectrometry, in vitro ubiquitination assays with lysine-less and N-terminally blocked substrates, N-terminal sequencing, active-site mutagenesis |
The Journal of biological chemistry |
High |
23696636
|
| 2013 |
UBE2W forms a noncovalent homodimer via residues in the β-sheet region and Loops 4 and 7 of the catalytic domain; mutation of two catalytic domain residues disrupts dimerization, but dimerization is not required for ubiquitin transfer activity; the C-terminal region of UBE2W adopts a noncanonical position distinct from all other known E2 structures. |
Biochemical dimerization assays, mutagenesis, structural characterization |
Cell biochemistry and biophysics |
Medium |
23709311
|
| 2014 |
Ube2w recognizes the backbone atoms of intrinsically disordered N-termini of substrates (not sequence-specific); its own C-terminus is partly disordered and flexible, and this flexibility is required for substrate binding and modification — point mutations in or removal of the flexible C-terminus inhibits substrate binding and N-terminal ubiquitination. |
NMR solution ensemble of full-length Ube2w, mutagenesis, in vitro ubiquitination assays, computational modeling |
Nature chemical biology |
High |
25436519
|
| 2015 |
TRIM5α employs Ube2W to anchor Lys63-linked polyubiquitin chains via Ube2W-catalyzed monoubiquitination of TRIM5α (targeting internal lysines 45 and 50, not the N-terminus which is αN-acetylated in cells); these anchored chains are then elongated by Ube2N/Ube2V2; this anchored K63-linked polyUb is required for TRIM5α-mediated restriction of viral reverse transcription. |
In vitro ubiquitination assays, cell-based E2 depletion, ubiquitin mutant studies, retroviral restriction assays |
The EMBO journal |
High |
26101372
|
| 2010 |
UBE2W interacts with the FANCL E3 ligase via FANCL's PHD domain, catalyzes monoubiquitination of FANCL's PHD domain in vitro, and promotes monoubiquitination of FANCD2 in cells in response to UV irradiation (but not MMC), indicating a DNA-damage pathway-specific regulatory role distinct from UBE2T/HHR6. |
Co-immunoprecipitation, in vitro ubiquitination assay, overexpression and knockdown in mammalian cells, domain mapping |
Molecules and cells |
Medium |
21229326
|
| 2015 |
Ube2w KO mice show postnatal lethality and defects in testis, thymus, and skin; proteomic analysis of presymptomatic testis shows preferential accumulation of intrinsically disordered proteins in the absence of UBE2W, supporting the model that UBE2W preferentially targets disordered polypeptides in vivo; UBE2W is ubiquitously expressed as a single isoform localized to the cytoplasm. |
Germline knockout mouse generation, proteomics/mass spectrometry, immunofluorescence for subcellular localization, histology |
The Journal of biological chemistry |
High |
26601958
|
| 2016 |
Genetic epistasis in chicken and human cells shows that Ube2w does not have an essential role in the DNA damage response, but that loss of Ube2w suppresses the DNA damage hypersensitivity of Rnf4-deficient cells, indicating that Ube2w and Rnf4 function in distinct pathways in response to DNA damage despite their functional interaction in vitro. |
Genetic epistasis (double-mutant/knockdown), DNA damage sensitivity assays in chicken and human cells |
Scientific reports |
Medium |
27185577
|
| 2017 |
Ube2w deficiency markedly decreases mutant huntingtin (mHTT) aggregate formation and increases soluble monomeric mHTT in cultured cells and in HdhQ200 knock-in mice, while reducing mHTT-induced cytotoxicity; this places Ube2w upstream of mHTT aggregation and implicates N-terminal ubiquitination of the disordered HTT N-terminus in promoting aggregation. |
Ube2w knockdown/KO in immortalized cells, primary neurons, and knock-in mouse model; filter trap aggregation assays; cell viability assays |
Neurobiology of disease |
Medium |
28986324
|
| 2021 |
UBE2W N-terminally ubiquitinates UCHL1 and UCHL5 at their N-termini; N-terminal ubiquitination of these deubiquitinases distinctly alters their DUB activity; antibodies selectively recognizing N-terminal diglycine remnants (not isopeptide-linked diglycine on lysine) were used to map endogenous UBE2W substrates by mass spectrometry proteomics. |
Monoclonal antibody development, antibody-crystal structure determination, mass spectrometry proteomics, DUB activity assays |
Nature communications |
High |
34326324
|
| 2023 |
UBE2W acts as a writer of aminoacylated lysine ubiquitination, a newly identified PTM in which ubiquitin is attached to the α-amine of aminoacylated lysine residues; over 2,000 such ubiquitination sites were identified on all 20 aminoacylated lysines in human cell lines, and this modification mediates rapid protein degradation. |
Genetic code expansion with unnatural amino acids as chemical probes, mass spectrometry, cell-based ubiquitination assays |
Nature structural & molecular biology |
Medium |
36593310
|
| 2022 |
UBE2W inhibits TNFα-induced NF-κB transcriptional activity by attenuating IκB and p65 phosphorylation and blocking nuclear entry of p65; UBE2W overexpression ameliorates DSS-induced colitis in mice with reduced pro-inflammatory cytokine expression. |
Dual-luciferase reporter assay, Western blot, immunofluorescence, mouse colitis model with UBE2W overexpression/knockdown |
Digestive diseases and sciences |
Low |
35314916
|
| 2026 |
UBE2W mediates K63-linked ubiquitination of p53, regulates p53 subcellular localization, and thereby impairs p53-mediated DNA damage repair and apoptotic pathways in pancreatic cancer cells. |
In vitro and in vivo functional experiments, ubiquitination assays, subcellular fractionation, knockdown/overexpression |
American journal of cancer research |
Low |
41657785
|