A Click Chemistry Approach Reveals the Chromatin-Dependent Histone H3K36 Deacylase Nature of SIRT7.

ABSTRACT: Using an engineered pyrrolysyl-tRNA synthetase mutant together with tRNA_CUA^Pyl, we have genetically encoded N^ε-(7-azidoheptanoyl)-l-lysine (AzHeK) by amber codon in Escherichia coli for recombinant expression of a number of AzHeK-containing histone H3 proteins. We assembled in vitro acyl-nucleosomes from these recombinant acyl-H3 histones. All these acyl-nucleosomes contained an azide functionality that allowed quick click labeling with a strained alkyne dye for in-gel fluorescence analysis. Using these acyl-nucleosomes as substrates and click labeling as a detection method, we systematically investigated chromatin deacylation activities of SIRT7, a class III NAD⁺-dependent histone deacylase with roles in aging and cancer biology. Besides confirm

SUBMITTER: Wang WW 

PROVIDER: S-EPMC6812484 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature