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The viral IE protein WSV051 was a potential SUMOylation substrate. WSV051 was SUMOylated only by UBC9 and the active form of SUMO, SUMO-GG. The 20-μl reaction system contained 20 mM HEPES (pH 7.5), 5 mM MgCl2, 10 mM ATP, 50 nM E1, 5 μM E2, 9 μM SUMO/SUMO-GG/mSUMO, and 2 μM WSV051. The reactions were stopped by the addition of SDS-PAGE sample buffer, and the reaction mixtures were boiled for 5 min and then analyzed by Western blotting using antibodies against the potential substrates.
