Dataset Information


A role of RPA in replication coupled nucleosome assembly

ABSTRACT: Chromatin replication requires tight coordination of nucleosome assembly machinery with DNA replication machinery. While significant progress has been made in characterizing histone chaperones in this process, the mechanism of whereby nucleosome assembly couples with DNA replication remains largely unknown. Here we show that replication protein A (RPA), a single-stranded DNA (ssDNA) binding protein that is essential for DNA replication provides a binding platform for H3-H4 deposition by histone chaperons and is required for nucleosome formation on nascent chromatin. RPA binds free histone H3-H4 but not nucleosomal histones, and a RPA coated ssDNA stimulates assembly of H3-H4 onto double strand DNA in vitro. RPA mutant with reduced H3-H4 binding exhibits synthetic genetic interaction with mutations at key factors involved in replication-coupled (RC) nucleosome assembly, and are defective in assembly of replicating DNA into nucleosomes in cells. These results reveal a novel function for RPA in nucleosome assembly and a mechanism whereby nucleosome assembly is coordinated with DNA replication. Overall design: To thoroughly compare nucleosome formation on nascent chromatin between wild type and RPA mutant cells, we mapped nucleosomes and genomic DNA in wild type, rfa1-A88P and rtt109∆ cells at G1 and early S phase. Two independent biological repeats were performed for each sample.

INSTRUMENT(S): Illumina HiSeq 2000 (Saccharomyces cerevisiae)


PROVIDER: GSE83648 | GEO | 2017-01-27



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RPA binds histone H3-H4 and functions in DNA replication-coupled nucleosome assembly.

Liu Shaofeng S   Xu Zhiyun Z   Leng He H   Zheng Pu P   Yang Jiayi J   Chen Kaifu K   Feng Jianxun J   Li Qing Q  

Science (New York, N.Y.) 20170101 6323

DNA replication-coupled nucleosome assembly is essential to maintain genome integrity and retain epigenetic information. Multiple involved histone chaperones have been identified, but how nucleosome assembly is coupled to DNA replication remains elusive. Here we show that replication protein A (RPA), an essential replisome component that binds single-stranded DNA, has a role in replication-coupled nucleosome assembly. RPA directly binds free H3-H4. Assays using a synthetic sequence that mimics f  ...[more]

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