Decoding the protein composition of whole nucleosomes with Nuc-MS.

Schachner, Luis F; Jooß, Kevin; Morgan, Marc A; Piunti, Andrea; Meiners, Matthew J; Kafader, Jared O; Lee, Alexander S; Iwanaszko, Marta et al. · Nat Methods · 2021

basic_science · Level V

Where this comes from

Abstract

Current proteomic approaches disassemble and digest nucleosome particles, blurring readouts of the 'histone code'. To preserve nucleosome-level information, we developed Nuc-MS, which displays the landscape of histone variants and their post-translational modifications (PTMs) in a single mass spectrum. Combined with immunoprecipitation, Nuc-MS quantified nucleosome co-occupancy of histone H3.3 with variant H2A.Z (sixfold over bulk) and the co-occurrence of oncogenic H3.3K27M with euchromatic marks (for example, a >15-fold enrichment of dimethylated H3K79me2). Nuc-MS is highly concordant with chromatin immunoprecipitation-sequencing (ChIP-seq) and offers a new readout of nucleosome-level biology.

Medical subject headings