Intrinsic flexibility of ubiquitin on proliferating cell nuclear antigen (PCNA) in translesion synthesis.

Ubiquitin conjugation provides a crucial signalling role in hundreds of cellular pathways, however, a structural understanding of ubiquitinated substrates is lacking. One important substrate is monoubiquitinated PCNA (PCNA-Ub), which signals for recruitment of damage-tolerant polymerases in the translesion synthesis (TLS) pathway of DNA damage avoidance. We use a novel and ...
efficient enzymatic method to produce PCNA-Ub at high yield with a native isopeptide bond, and study its Usp1/UAF1 dependent deconjugation. In solution we find that the ubiquitin moiety is flexible relative to the PCNA, with its hydrophobic patch mostly accessible for recruitment of TLS polymerases, which promotes the interaction with polymerase η. The studies are a prototype for the nature of the ubiquitin modification.
J. Biol. Chem.
Date: Sep. 18, 2012
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