Detection of Histone Modification Dynamics during the Cell Cycle by MS-Based Proteomics
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).
Key wordsSILAC labeling PTM-analysis Mass spectrometry Histone quantitation
This work was supported by a grant from the Deutsche Forschungsgemeinschaft (DFG, CRC1064-A16).
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