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

Luis F SchachnerKevin JooßMarc Alard Jonathan MorganAndrea PiuntiMatthew J MeinersJared O KafaderAlexander S LeeMarta IwanaszkoMarcus A CheekJonathan M BurgSarah A HowardMichael-Christopher KeoghAli ShilatifardNeil L Kelleher
Published in: Nature methods (2021)
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.
Keyphrases
  • mass spectrometry
  • multiple sclerosis
  • ms ms
  • single cell
  • dna methylation
  • genome wide
  • transcription factor
  • gene expression
  • dna damage
  • copy number
  • healthcare
  • high throughput sequencing