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PMID: 19704001 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Protein phosphatase 2A-dependent dephosphorylation of replication protein A is required for the repair of DNA breaks induced by replication stress.

Molecular and cellular biology ·Vol. 29 ·No. 21 ·2009-11-00 ·Pages 5696-709

Feng J, Wakeman T, Yong S, Wu X, Kornbluth S, Wang XF

Abstract

Eukaryotic genomic integrity is safeguarded by cell cycle checkpoints and DNA repair pathways, collectively known as the DNA damage response, wherein replication protein A (RPA) is a key regulator playing multiple critical roles. The genotoxic insult-induced phosphorylation of the 32-kDa subunit of human RPA (RPA32), most notably the ATM/ATR-dependent phosphorylation at T21 and S33, acts to suppress DNA replication and recruit other checkpoint/repair proteins to the DNA lesions. It is not clear, however, how the DNA damage-responsive function of phosphorylated RPA is attenuated and how the replication-associated activity of the unphosphorylated form of RPA is restored when cells start to resume the normal cell cycle. We report here that in cells recovering from hydroxyurea (HU)-induced genotoxic stress, RPA32 is dephosphorylated by the serine/threonine protein phosphatase 2A (PP2A). Interference with PP2A catalytic activity causes persistent RPA32 phosphorylation and increased HU sensitivity. The PP2A catalytic subunit binds to RPA following DNA damage and can dephosphorylate RPA32 in vitro. Cells expressing a RPA32 persistent phosphorylation mimetic exhibit normal checkpoint activation and reenter the cell cycle normally after recovery but display a pronounced defect in the repair of DNA breaks. These data indicate that PP2A-mediated RPA32 dephosphorylation is required for the efficient DNA damage repair.

MeSH Terms
Cell Line DNA Damage DNA Repair/drug effects,radiation effects DNA Replication/drug effects,radiation effects Humans Hydroxyurea/pharmacology Mitosis/drug effects,radiation effects Phosphorylation/drug effects,radiation effects Protein Phosphatase 2/metabolism Replication Protein A/metabolism Serine/metabolism Stress, Physiological/drug effects,radiation effects Threonine/metabolism Ultraviolet Rays
Chemicals
Replication Protein A Threonine Serine RPA2 protein, human Protein Phosphatase 2 Hydroxyurea
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Feng Junjie
Department of Pharmacology and Cancer Biology, Duke University Medical Center, La Salle Street Extension, Durham, NC 27710, USA.
Wakeman Timothy
Yong Sheila
Wu Xiaohua
Kornbluth Sally
Wang Xiao-Fan
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
1098-5549
DOI
10.1128/MCB.00191-09
Published
2009-11-00
Epub
2009-00-24
Pages
5696-709
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC2772729
Subset
IM
Grants
NCI NIH HHS · R01 CA123250 · United States
NCI NIH HHS · CA123250 · United States
External Links
PubMed source DOI: 10.1128/MCB.00191-09 PMC Full Text View journal details
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