Figure 1.
The H4 basic patch is required for proper H2BK123 H2Bub1. A, sequence of the H4 N terminus with basic patch residues depicted. Arrows show the arginine residues mutated to alanine to create the basic patch mutant H42RA used throughout the study. B, immunoblotting was performed for the indicated proteins or modifications using extracts from cells that expressed WT H4 (H4WT) or the basic patch mutant (H42RA). Strains that had a mutation at H3 lysine 79 (H3K79R) or were deleted for the E3 ubiquitin ligase Bre1 (bre1Δ) were used as antibody controls for H3K79 methylation and H2Bub1, respectively. H3 was used as a loading control. C, representative immunoblot analysis of H4WT and H42RA at increasing concentrations for H2B or H2Bub1. D, relative quantification of H2Bub1/H2B from C. The means ± S.E. were analyzed from three biological replicates.