ATAD5 promotes replication restart by regulating RAD51 and PCNA in response to replication stress

Nat Commun. 2019 Dec 16;10(1):5718. doi: 10.1038/s41467-019-13667-4.

Abstract

Maintaining stability of replication forks is important for genomic integrity. However, it is not clear how replisome proteins contribute to fork stability under replication stress. Here, we report that ATAD5, a PCNA unloader, plays multiple functions at stalled forks including promoting its restart. ATAD5 depletion increases genomic instability upon hydroxyurea treatment in cultured cells and mice. ATAD5 recruits RAD51 to stalled forks in an ATR kinase-dependent manner by hydroxyurea-enhanced protein-protein interactions and timely removes PCNA from stalled forks for RAD51 recruitment. Consistent with the role of RAD51 in fork regression, ATAD5 depletion inhibits slowdown of fork progression and native 5-bromo-2'-deoxyuridine signal induced by hydroxyurea. Single-molecule FRET showed that PCNA itself acts as a mechanical barrier to fork regression. Consequently, DNA breaks required for fork restart are reduced by ATAD5 depletion. Collectively, our results suggest an important role of ATAD5 in maintaining genome integrity during replication stress.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATPases Associated with Diverse Cellular Activities / genetics
  • ATPases Associated with Diverse Cellular Activities / metabolism*
  • Bromodeoxyuridine / metabolism
  • Cell Line, Tumor
  • DNA Breaks / drug effects
  • DNA Repair
  • DNA Replication / drug effects
  • DNA Replication / genetics*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Flow Cytometry
  • Fluorescence Resonance Energy Transfer
  • Gene Knockdown Techniques
  • Genomic Instability / drug effects
  • Genomic Instability / genetics*
  • HEK293 Cells
  • Humans
  • Hydroxyurea / pharmacology
  • Proliferating Cell Nuclear Antigen / metabolism*
  • Protein Binding / drug effects
  • RNA, Small Interfering / metabolism
  • Rad51 Recombinase / metabolism*
  • Single Molecule Imaging

Substances

  • ATAD5 protein, human
  • DNA-Binding Proteins
  • PCNA protein, human
  • Proliferating Cell Nuclear Antigen
  • RNA, Small Interfering
  • RAD51 protein, human
  • Rad51 Recombinase
  • ATPases Associated with Diverse Cellular Activities
  • Bromodeoxyuridine
  • Hydroxyurea