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Highlights • ∼25% of human E2s have a ubiquitination site (Lys+5) near the catalytic center • Flexibility of the active-site region enables Lys+5 autoubiquitination in UBE2S • Lys+5-linked ubiquitin inhibits UBE2S by adopting a closed conformation • The extent of Lys+5 ubiquitination in UBE2S is regulated during the cell cycle Summary Ubiquitin-conjugating enzymes (E2s) govern key aspects of ubiquitin signaling.
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