David Cortez
Faculty Member
Last active: 2/4/2016

ATRIP binding to replication protein A-single-stranded DNA promotes ATR-ATRIP localization but is dispensable for Chk1 phosphorylation.

Ball HL, Myers JS, Cortez D
Mol Biol Cell. 2005 16 (5): 2372-81

PMID: 15743907 · PMCID: PMC1087242 · DOI:10.1091/mbc.e04-11-1006

ATR associates with the regulatory protein ATRIP that has been proposed to localize ATR to sites of DNA damage through an interaction with single-stranded DNA (ssDNA) coated with replication protein A (RPA). We tested this hypothesis and found that ATRIP is required for ATR accumulation at intranuclear foci induced by DNA damage. A domain at the N terminus of ATRIP is necessary and sufficient for interaction with RPA-ssDNA. Deletion of the ssDNA-RPA interaction domain of ATRIP greatly diminished accumulation of ATRIP into foci. However, the ATRIP-RPA-ssDNA interaction is not sufficient for ATRIP recognition of DNA damage. A splice variant of ATRIP that cannot bind to ATR revealed that ATR association is also essential for proper ATRIP localization. Furthermore, the ATRIP-RPA-ssDNA interaction is not absolutely essential for ATR activation because ATR phosphorylates Chk1 in cells expressing only a mutant of ATRIP that does not bind to RPA-ssDNA. These data suggest that binding to RPA-ssDNA is not the essential function of ATRIP in ATR-dependent checkpoint signaling and ATR has an important function in properly localizing the ATR-ATRIP complex.

MeSH Terms (25)

Adaptor Proteins, Signal Transducing Ataxia Telangiectasia Mutated Proteins Base Sequence Binding Sites Cell Cycle Proteins Cell Line Cell Nucleus Checkpoint Kinase 1 DNA, Single-Stranded DNA-Binding Proteins DNA Damage Exodeoxyribonucleases Humans In Vitro Techniques Phosphoproteins Phosphorylation Protein-Serine-Threonine Kinases Protein Binding Protein Kinases Protein Structure, Tertiary Replication Protein A RNA, Small Interfering Signal Transduction Transfection Two-Hybrid System Techniques

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