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WASp modulates RPA function on single-stranded DNA in response to replication stress and DNA damage.

Seong-Su Han1,2, Kuo-Kuang Wen1,2, María L García-Rubio3

  • 1Department of Pediatrics, PennState College of Medicine, PennState Health Children's Hospital, Hershey, PA, 17033, USA.

Nature Communications
|June 29, 2022
PubMed
Summary
This summary is machine-generated.

Wiskott-Aldrich syndrome protein (WASp) is crucial for maintaining genomic stability by ensuring proper DNA replication fork function and DNA damage response. WASp deficiency destabilizes key protein complexes, leading to genomic instability.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Genomic instability, a hallmark of Wiskott-Aldrich syndrome (WAS), arises from perturbations in the replication-stress response (RSR) and DNA-damage response (DDR).
  • The precise mechanisms underlying genomic instability in WAS remain largely uncharacterized.
  • Replication protein A (RPA), a critical single-strand DNA (ssDNA) binding protein, plays vital roles in both RSR and DDR.

Purpose of the Study:

  • To elucidate the role of Wiskott-Aldrich syndrome protein (WASp) in modulating RPA function at perturbed replication forks.
  • To investigate the impact of WASp deficiency on DNA replication and repair pathways.
  • To understand the evolutionary conservation of WASp's function in DNA stress response.

Main Methods:

  • Investigated the localization and function of WASp at replication forks following genotoxic insult in human lymphocytes.
  • Assessed the impact of WASp deficiency on the accumulation of key DDR proteins (RPA, ATR, ETAA1, TOPBP1) and CHK1 activation.
  • Utilized yeast (Saccharomyces cerevisiae) models deficient in the WASp homolog (Las17) to study DNA recombination and double-strand break repair dynamics.

Main Results:

  • WASp accumulates at replication forks upon genotoxic insult, associates with RPA, and promotes RPA:ssDNA complex formation.
  • WASp deficiency leads to destabilized RPA:ssDNA complexes, impaired recruitment of DDR factors to perturbed replication forks, and reduced CHK1 activation.
  • Yeast cells lacking Las17 exhibit decreased RPA accumulation, impaired DNA recombination, and increased DNA double-strand break (DSB)-induced single-strand annealing (SSA), resulting in higher DSB frequencies.

Conclusions:

  • WASp plays an essential, evolutionarily conserved role in the DNA stress-resolution pathway by regulating RPA function at replication forks.
  • WASp deficiency results in RPA dysfunction, leading to global replication fork dysfunction and increased genomic instability.
  • These findings reveal a novel mechanism linking WASp to the maintenance of genomic integrity, crucial for understanding Wiskott-Aldrich syndrome pathogenesis.