Abstract
In dividing cells, faithful duplication of the genome must be accompanied by reproduction of the chromatin landscape on new DNA. Our research focuses on how human cells replicate chromatin and ensure transmission of genetic and epigenetic information during cell division.Recently, we have developed an explorative proteomic approach, Nascent Chromatin Capture (NCC), to profile chromatin replication in human cells. NCC relies on biotin‐dUTP labeling of replicating DNA and cross‐linking, followed by affinity‐purification of tagged chromatin and quantitative proteomics. By comparing nascent newly synthesized chromatin with mature post‐replicative chromatin, we provide association dynamics for 3995 proteins. Highly enriched in nascent chromatin, we find the complete human replisome followed by central players in chromatin assembly and cohesion establishment. In contrast, a large set of chromatin proteins including histone H1 show delayed association. By combining NCC enrichment with experimentally derived chromatin probabilities, we also use the data set to identify new replication factors. Together, this provides an extensive resource to understand genome and epigenome maintenance. In addition, this proteomic instrument provides opportunity to address transmission of histone marks and replication stress responses.
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