Investigating the cell cycle-associated dynamics of histone modifications using quantitative mass spectrometry.
In eukaryotic cells, posttranslational modifications (PTMs) on histones regulate chromatin structure and thus impact nearly all chromatin-templated events, including replication, transcription, and DNA repair. During S phase, newly synthesized histones are deposited onto DNA, leading to dilution of total chromatin-associated modifications. To maintain genome organization in daughter cells, histone PTMs must be reestablished in the subsequent cell cycle. Owing to their importance for determining cellular fate, the mechanisms that underlie the inheritance of epigenetic mark from parent cells by daughter cells are of great interest. In recent years, mass spectrometry (MS) has emerged as a powerful tool for identifying and quantifying histone modifications. This chapter describes strategies for investigating the reestablishment of histone PTMs during the mitotic cell cycle using quantitative MS approaches. By introducing these basic principles of experimental design and common protocols, we hope that this chapter will help readers to apply quantitative MS in their own research systems to study the biology of histone modifications.
National Institute of Biological Sciences, Beijing, PR China.
Mo
M
Xu
She
S
Chen
Bing
B
Zhu
D000478:Alkylation; D000596:Amino Acids / Q000737:chemistry / Q000378:metabolism; D000812:Anhydrides / Q000737:chemistry; D018929:Cell Culture Techniques; D002453:Cell Cycle; D059447:Cell Cycle Checkpoints; D056148:Chromatography, Reverse-Phase; D006367:HeLa Cells; D006657:Histones / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007553:Isotope Labeling; D010446:Peptide Fragments / Q000737:chemistry; D010449:Peptide Mapping; D011422:Propionates / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D059748:Proteolysis; D012015:Reference Standards; D053719:Tandem Mass Spectrometry / Q000592:standards; D014357:Trypsin / Q000737:chemistry
D000596:Amino Acids; D000812:Anhydrides; D006657:Histones; D010446:Peptide Fragments; D011422:Propionates; C096126:propionic anhydride; D014357:Trypsin