Loss of Dot1 abolishes histone H3 Lys 79 methylation
in vivo. (A) Western blots of yeast whole-cell extracts and
recombinant yeast histone H3 were probed with either antibodies against
the histone H3 tail (residues 1–20) or against a peptide containing
methylated Lys 79. (B) Specificity of the anti-methyl Lys 79
antibody. Antibodies (1/10,000 dilution) were preincubated with either
no peptide, 25 μg nonmethylated peptide, or 25 μg of methylated Lys
79 peptide for 30 min at room temperature before probing yeast
whole-cell extracts for histone H3. (C) Western blot screen
for histone H3 Lys 79 methylase. Whole-cell extracts from the indicated
deletion strains were probed with antibodies against methylated Lys 4,
methylated Lys 79, and unmodified H3. (D) Restoration of Lys
79 methylation by constructs containing the DOT1 gene.
Whole-cell extracts from the indicated strains were probed with either
anti-methyl Lys 4, or anti-methyl Lys 79 antibodies.