Gcn5 forms complexes with CRL4Cdt2 E3 ligase and And-1.
A, Cdt2, DDB1, Cul4A, and And-1 interact with full-length (Full) Gcn5 and Gcn5 mutant (492–837). 293T cells transfected with the indicated FLAG-Gcn5 plasmids were harvested 36 h after transfection for immunoprecipitation using anti-FLAG M2-agarose beads. The FLAG immunoprecipitates (IP) were then subjected to immunoblot (IB) using anti-Cdt2, anti-Cul4A, anti-DDB1, anti-And-1, anti-actin, or anti-FLAG antibody. B, And-1 does not interact with Cdt2. 293T cells transfected with the indicated plasmids were treated as in A. Note that Gcn5 was detected in FLAG-And-1 precipitates.