Detection of Histone Modification Dynamics during the Cell Cycle by MS-Based Proteomics

Methods Mol Biol. 2018:1832:61-74. doi: 10.1007/978-1-4939-8663-7_4.

Abstract

DNA replication and subsequent deposition of nucleosomes is critical for the maintenance of the genome and epigenetic inheritance. Experiments using human tissue culture cells harvested at defined stages of the cell cycle can help to elucidate the mechanism of histone deposition and chromatin assembly in detail. Here, we describe a pulsed-SILAC approach to distinguish newly synthesized and deposited histones during S-phase of the cell cycle from parental "old" histones incorporated in previous replications and to decipher posttranslational histone modifications (PTMs).

Keywords: Histone quantitation; Mass spectrometry; PTM-analysis; SILAC labeling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acylation
  • Cell Cycle*
  • Flow Cytometry
  • HEK293 Cells
  • HeLa Cells
  • Histones / metabolism*
  • Humans
  • Isotope Labeling
  • Peptides / metabolism
  • Protein Processing, Post-Translational*
  • Proteomics / methods*
  • Solubility
  • Sulfuric Acids / chemistry
  • Tissue Culture Techniques

Substances

  • Histones
  • Peptides
  • Sulfuric Acids
  • sulfuric acid