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TRIM5-21R proteins can be autoubiquitylated in vitro. Purified monomeric (lanes 1 to 6) and dimeric (lanes 7 to 12) TRIM5-21R proteins were incubated with the wheat Uba1 E1 enzyme, ubiquitin, and one of three ubiquitin E2-conjugating enzymes—UbcH5c, UbcH6, or UbcH7—in the presence or absence of ATP. Reactions were terminated after 60 min, and TRIM5-21R reaction products were analyzed by Western blotting (α-FLAG).
