A role of RPA in replication coupled nucleosome assembly

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.

Dataset features

Information

Specifications

Dataset type:	Genome binding/occupancy profiling by high throughput sequencing, Epigenomics
Number of samples:	40
Release date:	Jan 27 2017
Last update date:	Oct 16 2017
Access:	Public
Genes:	RPA1
Dataset link	A role of RPA in replication coupled nucleosome assembly

Experimental Protocol

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.

Repositories

GEO

GSE83648

ENA

SRP076987

BioProject

PRJNA326595

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Contact

Qing Li