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

Science. 2017 Jan 27;355(6323):415-420. doi: 10.1126/science.aah4712.

Abstract

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 freshly unwound single-stranded DNA at replication fork showed that RPA promotes DNA-(H3-H4) complex formation immediately adjacent to double-stranded DNA. Further, an RPA mutant defective in H3-H4 binding exhibited attenuated nucleosome assembly on nascent chromatin. Thus, we propose that RPA functions as a platform for targeting histone deposition to replication fork, through which RPA couples nucleosome assembly with ongoing DNA replication.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatin Assembly and Disassembly*
  • DNA Replication*
  • DNA, Single-Stranded / metabolism
  • Electrophoretic Mobility Shift Assay
  • Histone Chaperones / metabolism
  • Histones / metabolism*
  • Nucleosomes / metabolism*
  • RNA Polymerase I / genetics
  • RNA Polymerase I / metabolism*
  • Replication Protein A / genetics
  • Replication Protein A / metabolism*
  • Saccharomyces cerevisiae
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*

Substances

  • DNA, Single-Stranded
  • Histone Chaperones
  • Histones
  • Nucleosomes
  • RFA1 protein, S cerevisiae
  • RFA2 protein, S cerevisiae
  • RPA14 protein, S cerevisiae
  • Replication Protein A
  • Saccharomyces cerevisiae Proteins
  • RNA Polymerase I