期刊
MOLECULAR CELL
卷 65, 期 1, 页码 117-130出版社
CELL PRESS
DOI: 10.1016/j.molcel.2016.11.016
关键词
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资金
- Francis Crick Institute
- Cancer Research UK [FC001066]
- UK Medical Research Council [FC001066]
- Wellcome Trust [FC001066]
- Wellcome Trust Senior Investigator Award [106252/Z/14/Z]
- European Research Council Advanced Grant [669424-CHROMOREP]
- European Commission Marie Curie fellowship (FP7-PEOPLE-IEF)
- Federation of European Biochemical Societies Return-to-Europe fellowship
- Wellcome Trust [106252/Z/14/Z] Funding Source: Wellcome Trust
- Cancer Research UK [15669] Funding Source: researchfish
- The Francis Crick Institute [10397, 10349, 10002, 10068] Funding Source: researchfish
- The Francis Crick Institute
- Cancer Research UK [10066] Funding Source: researchfish
- Wellcome Trust [106252/Z/14/Z] Funding Source: researchfish
The integrity of eukaryotic genomes requires rapid and regulated chromatin replication. How this is accomplished is still poorly understood. Using purified yeast replication proteins and fully chromatinized templates, we have reconstituted this process in vitro. We show that chromatin enforces DNA replication origin specificity by preventing non-specific MCM helicase loading. Helicase activation occurs efficiently in the context of chromatin, but subsequent replisome progression requires the histone chaperone FACT (facilitates chromatin transcription). The FACT-associated Nhp6 protein, the nucleosome remodelers INO80 or ISW1A, and the lysine acetyltransferases Gcn5 and Esa1 each contribute separately to maximum DNA synthesis rates. Chromatin promotes the regular priming of lagging-strand DNA synthesis by facilitating DNA polymerase alpha function at replication forks. Finally, nucleosomes disrupted during replication are efficiently re-assembled into regular arrays on nascent DNA. Our work defines the minimum requirements for chromatin replication in vitro and shows how multiple chromatin factors might modulate replication fork rates in vivo.
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