FIG. 2.

In vitro acetylation of GATA-1 by CBP-HAT. (Top) GATA-1 constructs used as substrates for CBP-HAT. All recombinant proteins were expressed as GST fusion proteins. GATA-1, full-length GATA-1; GATA-1Δf, GATA-1 lacking the zinc finger region and the lysine-rich motifs (aa 198 to 316 deleted); f(GATA-1), the zinc finger region of GATA-1 including the lysine-rich motifs (aa 177 to 333); N-mut, K-to-A substitutions in the N-terminal lysine-rich motif in the context of full-length GATA-1; C-mut, K-to-A substitutions at the C-terminal motif; NC-mut, both motifs mutated. (Bottom) Acetylation of GATA-1 constructs by the HAT domain of CBP (CBP-HAT aa 1196 to 1718). The high-molecular-weight bands present in all lanes represent autoacetylated CBP-HAT. p/CAF-HAT, the HAT domain of p/CAF from aa 352 to 832.