Cited by 81DNA Repair MechanismsDNA and Nucleic Acid ChemistryEffects of Radiation Exposure

Preferential localization of hyperphosphorylated replication protein A to double-strand break repair and checkpoint complexes upon DNA damage

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Wu, Xiaoming, Yang, Zhengguan, Liu, Yiyong, Zou, Yue·2005·Biochemical Journal

Abstract

RPA (replication protein A) is an essential factor for DNA DSB (double-strand break) repair and cell cycle checkpoint activation. The 32 kDa subunit of RPA undergoes hyperphosphorylation in response to cellular genotoxic insults. However, the potential involvement of hyperphosphorylated RPA in DSB repair and checkpoint activation remains unclear. Using co-immunoprecipitation assays, we showed that cellular interaction of RPA with two DSB repair factors, Rad51 and Rad52, was predominantly mediated by the hyperphosphorylated species of RPA in cells after UV and camptothecin treatment. Moreover, Rad51 and Rad52 displayed higher affinity for the hyperphosphorylated RPA than native RPA in an in vitro binding assay. Checkpoint kinase ATR (ataxia telangiectasia mutated and Rad3-related) also interacted more efficiently with the hyperphosphorylated RPA than with native RPA following DNA damage. Consistently, immunofluorescence microscopy demonstrated that the hyperphosphorylated RPA was able to co-localize with Rad52 and ATR to form significant nuclear foci in cells. Our results suggest that hyperphosphorylated RPA is preferentially localized to DSB repair and the DNA damage checkpoint complexes in response to DNA damage.

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Authors
Wu, X., Yang, Z., Liu, Y., & Zou, Y.
Affiliations
East Tennessee State University (US)
Venue
Biochemical Journal
Published
2005-10-25
Volume
391
Issue
3
Pages
473-480
ISSN
  • 0264-6021 (ISSN-L)
  • 1470-8728
OpenAlex
W2063532739
Cited by
81
References
43
Citations by year
2025: 3 · 2024: 4 · 2023: 1 · 2022: 1 · 2021: 5 · 2020: 2 · 2019: 2 · 2017: 3 · 2016: 3 · 2015: 5 · 2014: 5 · 2013: 1 · 2012: 6
FWCI
2.26
Type
journal article
Language
en
Open access
Locations
  • Biochemical Journal (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Digital Commons - East Tennessee State University (East Tennessee State University) (submittedVersion)
Funders
  • East Tennessee State University
  • National Cancer Institute (R56CA086927, R01CA086927, CA-86927)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
  • Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Effects of Radiation Exposure
Keywords
replication protein A, DNA double-strand break repair, DNA damage checkpoint, ATR kinase, RAD51, RAD52, protein-protein interaction, UV-induced DNA damage, camptothecin, nuclear foci
MeSH terms
Adenocarcinoma, DNA Damage, DNA Repair, Gene Expression Regulation, Humans, Lung Neoplasms, Phosphorylation, Protein Binding, Ultraviolet Rays, Protein Transport, Cell Line, Tumor, Rad51 Recombinase, Replication Protein A, Rad52 DNA Repair and Recombination Protein

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