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Formation of Dynamic γ-H2AX Domains along Broken DNA Strands Is Distinctly Regulated by ATM and MDC1 and Dependent upon H2AX Densities in Chromatin
Open accessAbstract
A hallmark of the cellular response to DNA double-strand breaks (DSBs) is histone H2AX phosphorylation in chromatin to generate gamma-H2AX. Here, we demonstrate that gamma-H2AX densities increase transiently along DNA strands as they are broken and repaired in G1 phase cells. The region across which gamma-H2AX forms does not spread as DSBs persist; rather, gamma-H2AX densities equilibrate at distinct levels within a fixed distance from DNA ends. Although both ATM and DNA-PKcs generate gamma-H2AX, only ATM promotes gamma-H2AX formation to maximal distance and maintains gamma-H2AX densities. MDC1 is essential for gamma-H2AX formation at high densities near DSBs, but not for generation of gamma-H2AX over distal sequences. Reduced H2AX levels in chromatin impair the density, but not the distance, of gamma-H2AX formed. Our data suggest that H2AX fuels a gamma-H2AX self-reinforcing mechanism that retains MDC1 and activated ATM in chromatin near DSBs and promotes continued local phosphorylation of H2AX.
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“ATR and ATM also phosphorylate histone H2AX at Ser-139 (γH2AX) in response to DNA double-strand breaks (DSBs), which spreads along the DNA up to 200–400 kb and helps in the recruitment of proteins involved in DNA damage repair and checkpoint activation [29].”
From ATR- and ATM-Mediated DNA Damage Response Is Dependent on Excision Repair Assembly during G1 but Not in S Phase of Cell Cycle (2016) · cites this paper as [29]
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“Additionally, it is well established that ATR and ATM allow H2AX phosphorylation and spreading at the damage site, which changes the chromatin structure near the damage site and executes DNA repair through the HR pathway [21-23, 26, 63].”
From NER initiation factors, DDB2 and XPC, regulate UV radiation response by recruiting ATR and ATM kinases to DNA damage sites (2013) · cites this paper as [21]
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Fisher, M. R., Rivera-Reyes, A., Bloch, N. B., Schatz, D. G., & Bassing, C. H. (2017) · The Journal of Immunology · doi:10.4049/jimmunol.1601639 · Cited by 28
Self-citationRestoration of ATM Expression in DNA-PKcs–Deficient Cells Inhibits Signal End Joining
Neal, J. A., Xu, Y., Abe, M., Hendrickson, E. A., & Meek, K. (2016) · The Journal of Immunology · doi:10.4049/jimmunol.1501654 · Cited by 27
The nucleosome: orchestrating DNA damage signaling and repair within chromatin
Agarwal, P., & Miller, K. M. (2016) · Biochemistry and Cell Biology · doi:10.1139/bcb-2016-0017 · Cited by 28
A small ubiquitin binding domain inhibits ubiquitin-dependent protein recruitment to DNA repair foci
Helchowski, C. M., Skow, L., Roberts, K. H., Chute, C. L., & Canman, C. E. (2013) · Cell Cycle · doi:10.4161/cc.26640 · Cited by 28
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Barbieri, S., Babini, G., Morini, J., Friedland, W., Buonanno, M., Grilj, V., Brenner, D. J., Ottolenghi, A., & Baiocco, G. (2019) · Scientific Reports · doi:10.1038/s41598-019-50408-5 · Cited by 28
Leem, J., Kim, J., & Oh, J. S. (2023) · Nucleic Acids Research · doi:10.1093/nar/gkad213 · Cited by 25
Giorgio, E. D., Dalla, E., Tolotto, V., D’Este, F., Paluvai, H., Ranzino, L., & Brancolini, C. (2024) · Nucleic Acids Research · doi:10.1093/nar/gkae501 · Cited by 26
Garvin, A. J. (2019) · Biochemical Society Transactions · doi:10.1042/bst20190534 · Cited by 25
DNA Double Strand Break and Response Fluorescent Assays: Choices and Interpretation
Atkinson, J., Bezak, E., Le, H. V., & Kempson, I. M. (2024) · International Journal of Molecular Sciences · doi:10.3390/ijms25042227 · Cited by 25
Tubbs, A., Dorsett, Y., Chan, E., Helmink, B. A., Lee, B.-S., Hung, P. J., George, R., Bredemeyer, A. L., Mittal, A., Pappu, R. V., et al. (2014) · Molecular and Cellular Biology · doi:10.1128/mcb.00441-14 · Cited by 24
Self-citationSakai, H., Fujigaki, H., Mazur, S. J., & Appella, E. (2014) · Cell Cycle · doi:10.4161/cc.27920 · Cited by 24
Role of Polycomb Group Proteins in the DNA Damage Response – A Reassessment
Chandler, H., Patel, H., Palermo, R. D., Brookes, S., Matthews, N., & Peters, G. G. (2014) · PLoS ONE · doi:10.1371/journal.pone.0102968 · Cited by 22
Defective DNA damage response and repair in liver cells expressing hepatitis B virus surface antigen
Chung, Y. (2013) · The FASEB Journal · doi:10.1096/fj.12-226639 · Cited by 21
Schmalz, M. F., Liang, X., Wieser, I., Gruschel, C., Muskalla, L., Stöckl, M. T., Nitschke, R., Linz, N., Leitenstorfer, A., Vogel, A., et al. (2023) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.2220132120 · Cited by 20
Integrated signaling in developing lymphocytes
Bednarski, J. J., & Sleckman, B. P. (2012) · Cell Cycle · doi:10.4161/cc.22021 · Cited by 21
Self-citationPTEN enhances G2/M arrest in etoposide-treated MCF-7 cells through activation of the ATM pathway
ZHANG, R., Zhu, L., Zhang, L., Xu, A., Li, Z., Xu, Y., He, P. T., Wu, M., Wei, F., & WANG, C. (2016) · Oncology Reports · doi:10.3892/or.2016.4674 · Cited by 20
Retracted
Metadata
- Authors
- Savic, V., Yin, B., Maas, N. L., Bredemeyer, A. L., Carpenter, A. C., Helmink, B. A., Yang-Iott, K. S., Sleckman, B. P., & Bassing, C. H.
- Affiliations
- Children's Hospital of Philadelphia (US)
- Washington University in St. Louis (US)
- Corresponding author
- Bassing, Craig H.
- Venue
- Molecular Cell
- Publisher
- Elsevier BV
- Published
- 2009-05
- Volume
- 34
- Issue
- 3
- Pages
- 298-310
- ISSN
- 1097-2765 (ISSN-L)
- PMC
- PMC2744111
- OpenAlex
- W2036749550
- Cited by
- 204
- References
- 48
- Citations by year
- 2026: 1 · 2025: 7 · 2024: 14 · 2023: 9 · 2022: 11 · 2021: 12 · 2020: 14 · 2019: 12 · 2018: 7 · 2017: 12 · 2016: 9 · 2015: 9 · 2014: 12 · 2013: 22 · 2012: 15
- FWCI
- 7.12
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- www.elsevier.com/tdm/userlicense/1.0/ (tdm)
- www.elsevier.com/legal/tdmrep-license (tdm)
- www.elsevier.com/open-access/userlicense/1.0/ (vor, 1574 d)
- doi.org/10.15223/policy-017 (stm-asf)
- doi.org/10.15223/policy-037 (stm-asf)
- doi.org/10.15223/policy-012 (stm-asf)
- doi.org/10.15223/policy-029 (stm-asf)
- doi.org/10.15223/policy-004 (stm-asf)
- Locations
- Molecular Cell (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Figshare (submittedVersion)
- Funders
- National Cancer Institute (R01CA125195)
- National Institute of Allergy and Infectious Diseases (R01AI074953, T32AI055428)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Effects of Radiation Exposure
- Keywords
- γH2AX, DNA double-strand breaks, ATM, MDC1, DNA-PKcs, H2AX phosphorylation, DNA repair, G1 phase, chromatin
- MeSH terms
- Animals, B-Lymphocytes, Chromatin, DNA, DNA Damage, DNA-Binding Proteins, Endonucleases, Histones, Nuclear Proteins, Recombination, Genetic, Thymus Gland, Tumor Suppressor Protein p53, G1 Phase, Protein Serine-Threonine Kinases, Mice, Knockout, Cell Cycle Proteins, Genes, T-Cell Receptor alpha, Tumor Suppressor Proteins, Intracellular Signaling Peptides and Proteins, Adaptor Proteins, Signal Transducing, Mice, Ataxia Telangiectasia Mutated Proteins
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