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. 2009 Sep 22;37(21):7072–7084. doi: 10.1093/nar/gkp777

Figure 3.

Figure 3.

The C terminus of PCAF is responsible for the inhibition of cyclin A/cdk2 activity in vitro. (A) Schematic representation of the domains of PCAF used in the experiments. (B) Purified recombinant cyclin A/cdk2 complexes (400 nM) were incubated with increasing concentrations of Nterm, Cterm, Cterm ΔHAT and full-length PCAF and kinase assays were performed. (C) The same as in (B), but using different recombinant fragments of PCAF: HAT, ADA, Bromo and ADA-Bromo. (D) A typical kinase assay using purified recombinant cyclin A/cdk2 complex (400 nM) as a kinase and histone H1 (2 µg) as a substrate. Different reactions were performed by adding GST or different purified fragments of PCAF at a concentration of 800 nM. After incubation, reactions were stopped and samples were electrophoresed. Phosphorylation of histone H1 was detected by autoradiography using a PhosphorImager.