FIG. 3.

Acetylation increases MyoD affinity for CBP in vitro. (A) Equivalent amounts of wild-type (wt) and point-mutated (m2) MyoD were incubated with GST-CBP 1-2441-coated beads for 1 h, with or without acetyl-CoA (1 mM), and washed with TE buffer containing the indicated concentrations of KCl. GST-coated beads were used as a negative control. (B) Autoradiograms of the blots shown in panel A were analyzed by densitometry. Open circle, nonacetylated MyoD; solid square, acetylated MyoD. Increasing doses (2, 10, and 50 ng) of recombinant MyoD were incubated with GST-CBP absorbed onto glutathione-coated beads in the presence or absence of acetyl-CoA. The presence of MyoD in association with CBP was detected by Western blotting as described for Fig. 2.