RP - Large-scale in vitro PCNA ubiquitinylation by Rad18/Rad6 (»Add to Infobox)

Sixma Titia

Titia Sixma's Group

PCNA is monoubiquitinylated on lysine K164 by the E2/E3 enzymes Rad6/Rad18. This modification provides an essential first step in DNA damage tolerance pathways by signaling for recruitment of Y-family polymerases capable of replication past sites of DNA damage. We have reconstituted this ubiquitinylation reaction in vitro using recombinantly expressed E1, Rad6/Rad18, PCNA and ubiquitin proteins on a μM scale. The modified product is specifically ubiquitinylated on K164 and is suitable for biochemical experiments such as binding studies or activity assays for deubiquitinylation enzymes.

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Sixma Titia (Principal Investigator / Titia Sixma's Group)

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