학술논문
Decoding the protein composition of whole nucleosomes with Nuc-MS
nucleosome mass spectrometry
nucleosome mass spectrometry
Document Type
Report
Author
Source
Nature Methods. March 2021, Vol. 18 Issue 3, p303, 6 p.
Subject
Language
English
ISSN
1548-7091
Abstract
Author(s): Luis F. Schachner [sup.1] [sup.2] , Kevin Jooß [sup.1] [sup.3] , Marc A. Morgan [sup.4] [sup.5] , Andrea Piunti [sup.4] [sup.5] , Matthew J. Meiners [sup.6] , Jared O. [...]
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. Nuc-MS makes use of top-down mass spectrometry in 'native' mode to quantitatively interrogate histone proteoforms and their post-translational modifications in a single experiment.
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. Nuc-MS makes use of top-down mass spectrometry in 'native' mode to quantitatively interrogate histone proteoforms and their post-translational modifications in a single experiment.