Interactions of Human Mismatch Repair Proteins MutSα and MutLα with Proteins of the ATR-Chk1 Pathway
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At clinically relevant doses, chemotherapeutic S(N)1 DNA methylating agents induce an ATR-mediated checkpoint response in human cells that is dependent on functional MutSalpha and MutLalpha. Deficiency of either mismatch repair activity renders cells highly resistant to this class of drug, but the mechanisms linking mismatch repair to checkpoint activation have remained elusive. In this study we have systematically examined the interactions of human MutSalpha and MutLalpha with proteins of the ATR-Chk1 pathway using both nuclear extracts and purified proteins. Using nuclear co-immunoprecipitation, we have detected interaction of MutSalpha with ATR, TopBP1, Claspin, and Chk1 and interaction of MutLalpha with TopBP1 and Claspin. We were unable to detect interaction of MutSalpha or MutLalpha with Rad17, Rad9, or replication protein A in the extract system. Use of purified proteins confirmed direct interaction of MutSalpha with ATR, TopBP1, and Chk1 and of MutLalpha with TopBP1. MutSalpha-Claspin and MutLalpha-Claspin interactions were not demonstrable with purified proteins, suggesting that extract interactions are indirect or depend on post-translational modification. Use of a modified chromatin immunoprecipitation assay showed that proliferating cell nuclear antigen, ATR, TopBP1, and Chk1 are recruited to chromatin in a MutLalpha- and MutSalpha-dependent fashion after N-methyl-N'-nitro-N-nitrosoguanidine treatment. However, chromatin enrichment of replication protein A, Claspin, Rad17-RFC, and Rad9-Rad1-Hus1 was not detected in these experiments. Although our failure to observe enrichment of the latter activities could be due to sensitivity limitations, these observations may indicate a novel mechanism for ATR activation.
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The MMR pathway is required for ATR-Chk1 checkpoint activation.
“suggesting the MMR pathway is required for ATR-Chk1 checkpoint activation [214]”
From Functional interplay between ATM/ATR-mediated DNA damage response and DNA repair pathways in oxidative stress (2014) · cites this paper as [214]
MutSα associates with ATR, TopBP1, Claspin, and Chk1 but not with Rad17, Rad9, or RPA, while MutLα interacts only with TopBP1 and Claspin.
“MutSα associates with ATR, TopBP1, Claspin, and Chk1 (but not Rad17, Rad9, or RPA), whereas MutLα interacts only with TopBP1 and Claspin [214]”
From Functional interplay between ATM/ATR-mediated DNA damage response and DNA repair pathways in oxidative stress (2014) · cites this paper as [214]
MNNG-induced recruitment of ATR, TopBP1, and Chk1 to chromatin requires MutSα and MutLα.
“the MNNG-induced recruitment of ATR, TopBP1, and Chk1 to chromatin requires MutSα and MutLα”
From Functional interplay between ATM/ATR-mediated DNA damage response and DNA repair pathways in oxidative stress (2014) · cites this paper as [214]
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- Authors
- Liu, Y., Fang, Y., Shao, H., Lindsey-Boltz, L., Sancar, A., & Modrich, P.
- Affiliations
- Howard Hughes Medical Institute (US)
- University of North Carolina at Chapel Hill (US)
- Corresponding author
- Modrich, Paul
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2010-02
- Volume
- 285
- Issue
- 8
- Pages
- 5974-5982
- ISSN
- 0021-9258 (ISSN-L)
- PMC
- PMC2820822
- OpenAlex
- W2038663019
- Cited by
- 78
- References
- 61
- Citations by year
- 2026: 3 · 2025: 3 · 2024: 2 · 2022: 4 · 2021: 3 · 2020: 2 · 2019: 4 · 2018: 3 · 2017: 3 · 2016: 5 · 2015: 11 · 2014: 9 · 2013: 10 · 2012: 6
- FWCI
- 2.15
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- Carolina Digital Repository (University of North Carolina at Chapel Hill) (submittedVersion)
- UNC Libraries
- Funders
- Howard Hughes Medical Institute
- National Institutes of Health (GM45190)
- National Institute of General Medical Sciences (R01GM032833, GM 45190, GM32833, R01GM045190)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Health Sciences › Medicine › Pathology and Forensic Medicine › Genetic factors in colorectal cancer
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Keywords
- DNA mismatch repair, MutSα, MutLα, ATR-Chk1 pathway, DNA damage checkpoint, TopBP1, Claspin, Chk1, N-methyl-N'-nitro-N-nitrosoguanidine, protein-protein interaction, DNA replication checkpoint, chromatin recruitment
- MeSH terms
- MutL Proteins, Checkpoint Kinase 1, Carrier Proteins, DNA-Binding Proteins, HeLa Cells, Humans, Methylnitronitrosoguanidine, Nuclear Proteins, Protein Kinases, Protein Serine-Threonine Kinases, Cell Cycle Proteins, Proliferating Cell Nuclear Antigen, DNA Repair Enzymes, Adaptor Proteins, Signal Transducing, Replication Protein A, DNA Mismatch Repair, Ataxia Telangiectasia Mutated Proteins
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