Autoinhibition Mechanism of the Ubiquitin-Conjugating Enzyme UBE2S by Autoubiquitination.
Date
2019-08-06Author
Liess, AKL
Kucerova, A
Schweimer, K
Yu, L
Roumeliotis, TI
Diebold, M
Dybkov, O
Sotriffer, C
Urlaub, H
Choudhary, JS
Mansfeld, J
Lorenz, S
Type
Journal Article
Metadata
Show full item recordAbstract
Ubiquitin-conjugating enzymes (E2s) govern key aspects of ubiquitin signaling. Emerging evidence suggests that the activities of E2s are modulated by posttranslational modifications; the structural underpinnings, however, are largely unclear. Here, we unravel the structural basis and mechanistic consequences of a conserved autoubiquitination event near the catalytic center of E2s, using the human anaphase-promoting complex/cyclosome-associated UBE2S as a model system. Crystal structures we determined of the catalytic ubiquitin carrier protein domain combined with MD simulations reveal that the active-site region is malleable, which permits an adjacent ubiquitin acceptor site, Lys+5, to be ubiquitinated intramolecularly. We demonstrate by NMR that the Lys+5-linked ubiquitin inhibits UBE2S by obstructing its reloading with ubiquitin. By immunoprecipitation, quantitative mass spectrometry, and siRNA-and-rescue experiments we show that Lys+5 ubiquitination of UBE2S decreases during mitotic exit but does not influence proteasomal turnover of this E2. These findings suggest that UBE2S activity underlies inherent regulation during the cell cycle.
Collections
Subject
Cell Line
Hela Cells
Humans
Cysteine
Ubiquitin-Conjugating Enzymes
Lysine
Ubiquitin
Crystallography, X-Ray
Mitosis
Gene Expression Regulation
Catalytic Domain
Homeostasis
Ubiquitination
Molecular Dynamics Simulation
Research team
Post-translational modifications and cell proliferation
Language
eng
Date accepted
2019-05-17
License start date
2019-08
Citation
Structure (London, England : 1993), 2019, 27 (8), pp. 1195 - 1210.e7
Publisher
CELL PRESS