RPA Interacts with HIRA and Regulates H3.3 Deposition at Gene Regulatory Elements in Mammalian Cells

Honglian Zhang, Haiyun Gan, Zhiquan Wang, Jeong Heon Lee, Hui Zhou, Tamas Ordog, Marc S. Wold, Mats Ljungman, Zhiguo Zhang

Research output: Contribution to journalArticle

27 Scopus citations


The histone chaperone HIRA is involved in depositing histone variant H3.3 into distinct genic regions, including promoters, enhancers, and gene bodies. However, how HIRA deposits H3.3 to these regions remains elusive. Through a short hairpin RNA (shRNA) screening, we identified single-stranded DNA binding protein replication protein A (RPA) as a regulator of the deposition of newly synthesized H3.3 into chromatin. We show that RPA physically interacts with HIRA to form RPA-HIRA-H3.3 complexes, and it co-localizes with HIRA and H3.3 at gene promoters and enhancers. Depletion of RPA1, the largest subunit of the RPA complex, dramatically reduces both HIRA association with chromatin and the deposition of newly synthesized H3.3 at promoters and enhancers and leads to altered transcription at gene promoters. These results support a model whereby RPA, best known for its role in DNA replication and repair, recruits HIRA to promoters and enhancers and regulates deposition of newly synthesized H3.3 to these regulatory elements for gene regulation.

Original languageEnglish (US)
Pages (from-to)272-284
Number of pages13
JournalMolecular Cell
Issue number2
StatePublished - Jan 19 2017



  • enhancer
  • H3.3 deposition
  • HIRA
  • histone chaperone
  • nucleosome assembly
  • promoter
  • R-loop
  • RPA

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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