FIG. 9.
Acetylation of pre-RC proteins by Hbo1. Two micrograms of recombinant human Hbo1 protein was incubated with His6-geminin (lanes 2 and 5), His6-Cdc6 (lanes 3 and 6), GST-Orc2 (lanes 7 and 9), and His6-Mcm2 (lanes 8 and 10) in the presence of [3H]acetyl-CoA; the proteins were separated on 10% SDS-PAGE gels. The gels were stained with Coomassie (CBB) stain (lanes 4 to 6, 9, and 10), followed by fluorography (Fluorogram) (lanes 1 to 3, 7, and 8). GST protein (lanes 1 and 4) served as a negative control. Bands marked by asterisks identify labeled Hbo1 proteins produced by its autoacetylation.