2022 Annual Meeting
Investigating the Residue Specificity of p300 in Human Histone Peptides
We constructed a library of histone H3 mutants to identify the impact that the amino acid sequences around a lysine residue have on histone acetylation patterns. The first forty-five amino acids of histone H3, containing seven distinct lysine residues that are acetylated, were used for library construction. The library is comprised of seven subsets, wherein three amino acids flanking each lysine were randomized, while the six other lysines were mutated to arginine. A previously described yeast surface display based platform (Waldman et al) was used to visualize protein expression, enzyme expression, and acetylation levels. The library was sorted using flow cytometry to identify subpopulations with distinct acetylation levels, controlling for protein and enzyme expression. Subsequently, we used next generation DNA sequencing to identify amino acid sequence determinants of histone acetylation by p300 and identified links to protein targets of disease relevance.