Cited by 130DNA Repair MechanismsEffects of Radiation ExposureBacterial Genetics and Biotechnology

ATR and H2AX cooperate in maintaining genome stability under replication stress

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Chanoux, Rebecca A., Yin, Bu, Urtishak, Karen A., Asare, Amma, Bassing, Craig H., Brown, Eric J.·2009·Journal of Biological Chemistry

Abstract

Chromosomal abnormalities are frequently caused by problems encountered during DNA replication. Although the ATR-Chk1 pathway has previously been implicated in preventing the collapse of stalled replication forks into double-strand breaks (DSB), the importance of the response to fork collapse in ATR-deficient cells has not been well characterized. Herein, we demonstrate that, upon stalled replication, ATR deficiency leads to the phosphorylation of H2AX by ATM and DNA-PKcs and to the focal accumulation of Rad51, a marker of homologous recombination and fork restart. Because H2AX has been shown to play a facilitative role in homologous recombination, we hypothesized that H2AX participates in Rad51-mediated suppression of DSBs generated in the absence of ATR. Consistent with this model, increased Rad51 focal accumulation in ATR-deficient cells is largely dependent on H2AX, and dual deficiencies in ATR and H2AX lead to synergistic increases in chromatid breaks and translocations. Importantly, the ATM and DNA-PK phosphorylation site on H2AX (Ser(139)) is required for genome stabilization in the absence of ATR; therefore, phosphorylation of H2AX by ATM and DNA-PKcs plays a pivotal role in suppressing DSBs during DNA synthesis in instances of ATR pathway failure. These results imply that ATR-dependent fork stabilization and H2AX/ATM/DNA-PKcs-dependent restart pathways cooperatively suppress double-strand breaks as a layered response network when replication stalls.

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  1. Nghiem, P., Park, P. K., Kim, Y., Vaziri, C., and Schreiber, S. L. (2001) Proc. Natl. Acad. Sci. U. S. A. 98, 9092-9097
  2. Bassing, C. H., Chua, K. F., Sekiguchi, J., Suh, H., Whitlow, S. R., Fleming, J. C., Monroe, B. C., Ciccone, D. N., Yan, C., Vlasakova, K., Livingston, D. M., Ferguson, D. O., Scully, R., and Alt, F. W. (2002) Proc. Natl. Acad. Sci. U. S. A. 99, 8173-8178

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Authors
Chanoux, R. A., Yin, B., Urtishak, K. A., Asare, A., Bassing, C. H., & Brown, E. J.
Affiliations
  • University of Pennsylvania (US)
  • Children's Hospital of Philadelphia (US)
Corresponding author
Brown, Eric J.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2009-02
Volume
284
Issue
9
Pages
5994-6003
ISSN
0021-9258 (ISSN-L)
OpenAlex
W2091890593
Cited by
130
References
63
Citations by year
2026: 3 · 2025: 8 · 2024: 3 · 2023: 5 · 2022: 4 · 2021: 6 · 2020: 4 · 2019: 10 · 2018: 12 · 2017: 5 · 2016: 4 · 2015: 7 · 2014: 10 · 2013: 14 · 2012: 6
FWCI
3.98
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • Cancer Research Institute
  • Pennsylvania Department of Health
  • National Institutes of Health
  • National Institute on Aging (R01AG027376)
  • National Cancer Institute (R01CA125195)
  • Abramson Family Cancer Research Institute
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
  • Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Effects of Radiation Exposure
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
Keywords
ATR, H2AX, replication stress, genome stability, replication fork collapse, DNA double-strand breaks, ATM, DNA-PKcs, RAD51, homologous recombination, fork restart, chromatid breaks, translocation, checkpoint signaling
MeSH terms
Animals, Cells, Cultured, DNA Replication, DNA-Binding Proteins, Embryo, Mammalian, Fibroblasts, Histones, Metaphase, Mitosis, Nuclear Proteins, Phosphorylation, Radiation, Ionizing, RNA, Messenger, S Phase, Protein Serine-Threonine Kinases, Mice, Knockout, Cell Cycle Proteins, Reverse Transcriptase Polymerase Chain Reaction, Tumor Suppressor Proteins, Spectral Karyotyping, RNA, Small Interfering, Genomic Instability, Rad51 Recombinase, Mice, DNA-Activated Protein Kinase, DNA Breaks, Double-Stranded, Ataxia Telangiectasia Mutated Proteins

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