1. WASp modulates RPA function on single-stranded DNA in response to replication stress and DNA damage
- Author
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Seong-Su Han, Kuo-Kuang Wen, María L. García-Rubio, Marc S. Wold, Andrés Aguilera, Wojciech Niedzwiedz, Yatin M. Vyas, National Institutes of Health (US), National Institute of Allergy and Infectious Diseases (US), Cancer Research UK, European Research Council, Ministerio de Ciencia, Innovación y Universidades (España), Agencia Estatal de Investigación (España), and University of Iowa
- Subjects
DNA Replication ,Multidisciplinary ,Saccharomyces cerevisiae Proteins ,DNA Repair ,General Physics and Astronomy ,DNA, Single-Stranded ,General Chemistry ,Saccharomyces cerevisiae ,General Biochemistry, Genetics and Molecular Biology ,Genomic Instability ,DNA-Binding Proteins ,Replication Protein A ,Antigens, Surface ,Animals ,Humans ,DNA Breaks, Double-Stranded ,Wiskott-Aldrich Syndrome Protein ,Protein Binding - Abstract
Perturbation in the replication-stress response (RSR) and DNA-damage response (DDR) causes genomic instability. Genomic instability occurs in Wiskott-Aldrich syndrome (WAS), a primary immunodeficiency disorder, yet the mechanism remains largely uncharacterized. Replication protein A (RPA), a single-strand DNA (ssDNA) binding protein, has key roles in the RSR and DDR. Here we show that human WAS-protein (WASp) modulates RPA functions at perturbed replication forks (RFs). Following genotoxic insult, WASp accumulates at RFs, associates with RPA, and promotes RPA:ssDNA complexation. WASp deficiency in human lymphocytes destabilizes RPA:ssDNA-complexes, impairs accumulation of RPA, ATR, ETAA1, and TOPBP1 at genotoxin-perturbed RFs, decreases CHK1 activation, and provokes global RF dysfunction. las17 (yeast WAS-homolog)-deficient S. cerevisiae also show decreased ScRPA accumulation at perturbed RFs, impaired DNA recombination, and increased frequency of DNA double-strand break (DSB)-induced single-strand annealing (SSA). Consequently, WASp (or Las17)-deficient cells show increased frequency of DSBs upon genotoxic insult. Our study reveals an evolutionarily conserved, essential role of WASp in the DNA stress-resolution pathway, such that WASp deficiency provokes RPA dysfunction-coupled genomic instability., This work was supported in part by the NIH, National Institute of Allergy and Infectious Diseases (NIAID) grants R01AI146380 (to Y.M.V.), grant A24881 from ICR Intramural Grant and Cancer Research UK Programme (to W.N.), the European Research Council grant ERC2014 AdG669898 TARLOOP and the Spanish Ministry of Science and Innovation grant PDI2019-104270GB-I00/ BMC) (to A.A.), the University of Iowa Dance Marathon (UIDM) research award (to S.S.H), Research Bridge Award from the Carver College of Medicine University of Iowa (to Y.M.V), and the Endowments from the Mary Joy & Jerre Stead Foundation and from the PennState THON & Children’s Miracle Network (to Y.M.V). A subset of data was obtained at the Flow Cytometry Facility, which is a Carver College of Medicine/Holden Comprehensive Cancer Center core research facility at the University of Iowa. We thank the UIDM for supporting the research laboratory space where bulk of this work was carried out.
- Published
- 2022