Reconstitution of a human ATR-mediated checkpoint response to damaged DNA.
Open accessAbstract
The DNA damage checkpoint response delays cell cycle progression upon DNA damage and prevents genomic instability. Genetic analysis has identified sensor, mediator, signal transducer, and effector components of this global signal transduction pathway. Here we describe an in vitro system with purified human checkpoint proteins that recapitulates key elements of the DNA damage checkpoint. We show that the damage sensor ATR in the presence of topoisomerase II binding protein 1 (TopBP1) mediator/adaptor protein phosphorylates the Chk1 signal-transducing kinase in a reaction that is strongly dependent on the presence of DNA containing bulky base lesions. The dependence on damaged DNA requires DNA binding by TopBP1, and, indeed, TopBP1 shows preferential binding to damaged DNA. This in vitro system provides a useful platform for mechanistic studies of the human DNA damage checkpoint response.
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Chk1 is activated during HR following resection of DSBs and exposure of single-stranded DNA, via ATR kinase phosphorylation.
“Chk1 is activated during HR following the resection of DSBs and the exposure of single-stranded DNA. This elicits activation of the ATR kinase, which stimulates Chk1 activity via phosphorylation (Choi et al., 2007).”
From USP7 Is a Master Regulator of Genome Stability (2020) · cites this paper as Choi et al., 2007
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Self-citation
- Authors
- Choi, J.-H., Lindsey-Boltz, L. A., & Sancar, A.
- Affiliations
- University of North Carolina at Chapel Hill (US)
- Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, NC 27599-7260
- ORCID
- Sancar, Aziz
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2007-08-14
- Volume
- 104
- Issue
- 33
- Pages
- 13301-13306
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC1941640
- OpenAlex
- W1975767581
- Cited by
- 64
- References
- 35
- Citations by year
- 2026: 1 · 2025: 1 · 2024: 1 · 2023: 2 · 2022: 1 · 2020: 2 · 2018: 4 · 2017: 2 · 2016: 4 · 2015: 4 · 2014: 3 · 2013: 6 · 2012: 6
- FWCI
- 2.65
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Carolina Digital Repository (University of North Carolina at Chapel Hill) (submittedVersion)
- UNC Libraries
- Funders
- National Institutes of Health
- National Institute of General Medical Sciences (R01GM032833, GM32833)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Keywords
- DNA damage checkpoint, ATR kinase, TopBP1, Chk1, DNA damage sensor, signal transduction, DNA binding, cell cycle arrest, genomic instability, in vitro reconstitution
- MeSH terms
- Carrier Proteins, Cell-Free System, DNA, DNA Damage, DNA-Binding Proteins, HeLa Cells, Humans, Nuclear Proteins, Phosphorylation, Protein Binding, Protein Serine-Threonine Kinases, Cell Cycle Proteins, Ataxia Telangiectasia Mutated Proteins
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