Trim-Away degrades lysine-less substrates without requiring ligase autoubiquitination
Trim-Away degrades lysine-less substrates without requiring ligase autoubiquitination
ABSTRACT TRIM proteins are the largest family of E3 ligases in mammals. They include the intracellular antibody receptor TRIM21, which is responsible for mediating targeted protein degradation during Trim-Away. Despite their importance, the ubiquitination mechanism of TRIM ligases has remained elusive. Here we show that while TRIM21 activation results in ubiquitination of both ligase and substrate, autoubiquitination is regulatory and not required for substrate degradation. Substrate binding stimulates N-terminal RING autoubiquitination by the E2 Ube2W, but when inhibited by N-terminal acetylation this prevents neither substrate ubiquitination nor degradation and has no impact on TRIM21 antiviral activity. Instead, uncoupling ligase and substrate degradation prevents ligase recycling and extends functional persistence in cells. Substrate ubiquitination mediates degradation but Trim-Away efficiently degrades lysine-less substrates, suggesting a non-canonical ubiquitination mechanism explains its broad substrate specificity.
Kiss Leo、Luptak Jakub、Dickson Claire F、Rhinesmith Tyler、Weidenhausen Jonas、Maslen Sarah L、Sinning Irmgard、Clift Dean、Yang Ji-Chun、Neuhaus David、James Leo C、Smyly Shannon
MRC Laboratory of Molecular Biology, Francis Crick Avenue||Max Planck Institute of BiochemistryMRC Laboratory of Molecular Biology, Francis Crick AvenueEMBL Australia Node in Single Molecule Science and ARC Centre of Excellence in Advanced Molecular Imaging School of Medical Sciences, UNSW SydneyMRC Laboratory of Molecular Biology, Francis Crick AvenueBiochemiezentrum der Universit?t Heidelberg (BZH)||EMBL HeidelbergMRC Laboratory of Molecular Biology, Francis Crick Avenue||The Francis Crick InstituteBiochemiezentrum der Universit?t Heidelberg (BZH)MRC Laboratory of Molecular Biology, Francis Crick AvenueMRC Laboratory of Molecular Biology, Francis Crick AvenueMRC Laboratory of Molecular Biology, Francis Crick AvenueMRC Laboratory of Molecular Biology, Francis Crick AvenueMRC Laboratory of Molecular Biology, Francis Crick Avenue
分子生物学生物化学
Kiss Leo,Luptak Jakub,Dickson Claire F,Rhinesmith Tyler,Weidenhausen Jonas,Maslen Sarah L,Sinning Irmgard,Clift Dean,Yang Ji-Chun,Neuhaus David,James Leo C,Smyly Shannon.Trim-Away degrades lysine-less substrates without requiring ligase autoubiquitination[EB/OL].(2025-03-28)[2025-04-28].https://www.biorxiv.org/content/10.1101/2022.06.29.498105.点此复制
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