FIGURE 3.
NSL-associated MOF has a broader substrate specificity than MSL-associated MOF. A, acetylation of Hela long oligonucleosomes by MSL- and NSL-associated MOF. HAT assays were performed with HeLa long oligonucleosomes, [3H]AcCoA, and 1 μl (lane 2 and lane 4) or 2 μl (lane 3 and lane 5) of the indicated anti-FLAG (Fl) agarose eluates, and acetylated hisones were visualized by autoradiography. The MSL and NSL complexes used in these experiments contained approximately equal concentrations of MOF (Fig. 2A). B, acetylation of free recombinant histones by MSL- and NSL-associated MOF. HAT assays were performed with core histones, unlabeled acetyl CoA, and 0.6 μl (lanes 2 and 5), 1.2 μl (lanes 3 and 6), or 2.4 μl (lanes 4 and 7) of the indicated anti-FLAG agarose eluates, and modified residues of histone H4 were detected by Western blotting with acetylation-specific antibodies. C, acetylation of recombinant polynucleosomes by MSL- and NSL-associated MOF. HAT assays were performed with reconstituted polynucleosomes, unlabeled acetyl CoA, and 0.6 μl (lanes 2 and 5), 1.2 μl (lanes 3 and 6), or 2.4 μl (lanes 4 and 7) of the indicated anti-FLAG agarose eluates, and modified histone H4 residues were detected by Western blotting with acetylation-specific antibodies.