Investigating Mitotic Inheritance of Histone Posttranslational Modifications by Triple pSILAC Coupled to Nascent Chromatin Capture

Methods Mol Biol. 2022:2529:407-417. doi: 10.1007/978-1-0716-2481-4_17.

Abstract

Pulse stable isotope labeling with amino acids in cell culture (pSILAC) coupled to mass spectrometric analysis is a powerful tool to study propagation of histone post-translational modifications (PTMs). We describe the combination of triple pSILAC with pulse-chase labeling of newly replicated DNA by nascent chromatin capture (NCC). This technology tracks newly synthesized and recycled old histones, from deposition to transmission to daughter cells, unveiling principles of histone-based inheritance.

Keywords: Chromatin assembly; DNA replication; Histone; Mass spectrometry; Nascent Chromatin Capture; Posttranslational modifications; Pulse-chase; SILAC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatin* / genetics
  • Histone Code
  • Histones* / genetics
  • Histones* / metabolism
  • Mass Spectrometry
  • Protein Processing, Post-Translational

Substances

  • Chromatin
  • Histones