Proteome-wide identification of in vivo targets of DNA damage checkpoint kinases
Open accessAbstract
Understanding the role of DNA damage checkpoint kinases in the cellular response to genotoxic stress requires the knowledge of their substrates. Here, we report the use of quantitative phosphoproteomics to identify in vivo kinase substrates of the yeast DNA damage checkpoint kinases Mec1, Tel1, and Rad53 (orthologs of human ATR, ATM, and CHK2, respectively). By analyzing 2,689 phosphorylation sites in wild-type and various kinase-null cells, 62 phosphorylation sites from 55 proteins were found to be controlled by the DNA damage checkpoint. Examination of the dependency of each phosphorylation on Mec1 and Tel1 or Rad53, combined with sequence and biochemical analysis, revealed that many of the identified targets are likely direct substrates of these kinases. In addition to several known targets, 50 previously undescribed targets of the DNA damage checkpoint were identified, suggesting that a wide range of cellular processes is likely regulated by Mec1, Tel1, and Rad53.
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Effector kinases phosphorylate many targets, including cell cycle machinery and proteins important for replication and repair.
“Finally, effector kinases are responsible for the phosphorylation of a great number of targets, including cell cycle machinery factors and key proteins important for replication and repair [32,33].”
From Mind the gap: keeping UV lesions in check (2011) · cites this paper as [32]
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“Finally, effector kinases are responsible for the phosphorylation of a great number of targets, including cell cycle machinery factors and key proteins important for replication and repair [32,33].”
BackgroundKurat, C. F., Lambert, J.-P., van Dyk, D., Tsui, K., van Bakel, H., Kaluarachchi, S., Friesen, H., Kainth, P., Nislow, C., Figeys, D., et al. (2011) · Genes & Development · doi:10.1101/gad.173427.111 · Cited by 41 · see citing works
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- Authors
- Smolka, M. B., Albuquerque, C. P., Chen, S.-H., & Zhou, H.
- Affiliations
- Ludwig Cancer Research (US)
- University of California San Diego (US)
- Ludwig Cancer Research (BE)
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2007-06-19
- Volume
- 104
- Issue
- 25
- Pages
- 10364-10369
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC1965519
- OpenAlex
- W2107835389
- Cited by
- 404
- References
- 45
- Citations by year
- 2026: 2 · 2025: 8 · 2024: 14 · 2023: 12 · 2022: 21 · 2021: 15 · 2020: 10 · 2019: 17 · 2018: 26 · 2017: 19 · 2016: 26 · 2015: 24 · 2014: 23 · 2013: 25 · 2012: 35
- FWCI
- 12.73
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Ludwig Institute for Cancer Research
- National Human Genome Research Institute (K22HG002604)
- National Institute of General Medical Sciences (R01GM080469)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Keywords
- DNA damage checkpoint, checkpoint kinase, kinase substrates, Mec1, Tel1, Rad53, phosphorylation sites, quantitative phosphoproteomics, genotoxic stress response, Saccharomyces cerevisiae, ATR, ATM, Chk2
- MeSH terms
- Chromatography, Liquid, DNA Damage, Fungal Proteins, Methyl Methanesulfonate, Mutagens, Protein Kinases, Saccharomyces cerevisiae, Substrate Specificity, Protein Serine-Threonine Kinases, Cell Cycle Proteins, Proteome, Saccharomyces cerevisiae Proteins, Intracellular Signaling Peptides and Proteins, Tandem Mass Spectrometry, Checkpoint Kinase 2
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