Nascent chromatin capture proteomics determines chromatin dynamics during DNA replication and identifies unknown fork components (original) (raw)
“…In NCC, biotin-dUTP labeling of newly replicated DNA and cross-linking of associated proteins allow the isolation of nascent chromatin (Supplemental Fig. S1A,B; Alabert et al 2014). We combined NCC with pulsed SILAC (stable isotope labeling with amino acids in cell culture) to differentiate newly synthesized histones (heavy) and old recycled histones (light) ( Fig.…”
Section: Resultsmentioning
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“…In NCC, biotin-dUTP labeling of newly replicated DNA and cross-linking of associated proteins allow the isolation of nascent chromatin (Supplemental Fig. S1A,B; Alabert et al 2014). We combined NCC with pulsed SILAC (stable isotope labeling with amino acids in cell culture) to differentiate newly synthesized histones (heavy) and old recycled histones (light) ( Fig.…”
Section: Resultsmentioning
“…For an outline of the experimental design, see Supplemental Figure S7C. may require recruitment of enzyme cofactors (Alabert et al 2014;Kalb et al 2014). Such a tight regulation of PTM establishment could limit unwarranted silencing, which is jeopardized in cancers carrying EZH2-activating mutations (McCabe et al 2012).…”
Section: Cell Cycle Withdrawal Impacts Histone Ptm Levelsmentioning
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“…In mammals, some modifications are cast immediately after the replication fork by chromatin factors including chromatin remodelers and histone chaperones; other modifications are introduced later during chromatin maturation, when the replication fork has already passed. 8 In higher eukaryotes, genomic regions replicate asynchronously -transcriptionally active regions usually replicate earlier in the S-phase of cell cycle than do repressed heterochromatic regions. 9 Primarily, this replication schedule is defined by the pattern of active replication initiation sites (origins).…”
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“…For example, an extensive study in human cells revealed that in contrast to other histone modifications, H3K9me3 and H3K27me3 are inherited via the incorporation of the histones from the template chromatin (parental histones) to the nascent chromatin. 8 The mechanism that distinguishes H3K9me3 and H3K27me3 from the rest of the histone modifications during replication remains unknown.…”
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