A synthetic non-histone substrate to study substrate targeting by the Gcn5 HAT and sirtuin HDACs

Gcn5 and sirtuins are conserved HAT and HDAC enzymes that were first characterised as regulators of gene expression via histone acetylation. Although histone tails are important substrates of these enzymes, they also target non-histone proteins in diverse processes. Previously, we used SILAC-based mass spectrometry to identify novel non-histone substrates of Gcn5 and sirtuins in yeast. We found a shared target consensus sequence. In our latest work we used a synthetic biology approach to demonstrate that this consensus sequence can direct acetylation and deacetylation targeting by these enzymes in vivo. We used the synthetic substrate as a tool to prioritize SILAC-based acetylome analyses of SAGA mutants. Presented here are analyses of ada3∆ mutants analyzed in this work versus wild-type control. The strains and labels used are described in the BioRxiv report linked in the methods sections below.