Cell-cycle-dependent phosphorylation of the basal transcription factor RAP74

In this report, cell-cycle-dependent effects of TFIID on other basal transcription factors were investigated. We purified TFIID fractions from HeLa cells synchronized in the S/G2 phases and in early G1 phase, and show that RAP74 is phosphorylated more highly by the S/G2 phase TFIID fraction than by the early G1 phase TFIID fraction. Further analyses using deletion mutants of RAP74 revealed that amino acid residues 206-256 are phosphorylated by the TFIID fraction. Reconstitution of in vitro transcription activity indicates that the cell-cycle-dependent phosphorylation of RAP74 increases TFIIF transcription activity.