The DOI matches its Crossref record; 26 of the 26 references it cites were found in a registry or an index.
Quantitative Detection of125IdU-Induced DNA Double-Strand Breaks with γ-H2AX Antibody
Abstract
When mammalian cells are exposed to ionizing radiation and other agents that introduce DSBs into DNA, histone H2AX molecules in megabase chromatin regions adjacent to the breaks become phosphorylated within minutes on a specific serine residue. An antibody to this phosphoserine motif of human H2AX (gamma-H2AX) demonstrates that gamma-H2AX molecules appear in discrete nuclear foci. To establish the quantitative relationship between the number of these foci and the number of DSBs, we took advantage of the ability of (125)I, when incorporated into DNA, to generate one DNA DSB per radioactive disintegration. SF-268 and HT-1080 cell cultures were grown in the presence of (125)IdU and processed immunocytochemically to determine the number of gamma-H2AX foci. The numbers of (125)IdU disintegrations per cell were measured by exposing the same immunocytochemically processed samples to a radiation-sensitive screen with known standards. Under appropriate conditions, the data yielded a direct correlation between the number of (125)I decays and the number of foci per cell, consistent with the assumptions that each (125)I decay yields a DNA DSB and each DNA DSB yields a visible gamma-H2AX focus. Based on these findings, we conclude that gamma-H2AX antibody may form the basis of a sensitive quantitative method for the detection of DNA DSBs in eukaryotic cells.
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“Notably, the number of L1-induced DSBs is greater than the predicted numbers of successful insertions, suggesting a significant degree of inefficiency during the integration process.”
From The Human LINE-1 Retrotransposon Creates DNA Double-strand Breaks (2006) · cites this paper as [40]
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Cites
DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139
Rogakou, E. P., Pilch, D. R., Orr, A. H., Ivanova, V. S., & Bonner, W. M. (1998) · Journal of Biological Chemistry · doi:10.1074/jbc.273.10.5858 · Cited by 5,438
Self-citationMegabase Chromatin Domains Involved in DNA Double-Strand Breaks in Vivo
Rogakou, E. P., Boon, C., Redon, C., & Bonner, W. M. (1999) · The Journal of Cell Biology · doi:10.1083/jcb.146.5.905 · Cited by 2,422
Self-citationA critical role for histone H2AX in recruitment of repair factors to nuclear foci after DNA damage
Paull, T. T., Rogakou, E. P., Yamazaki, V., Kirchgessner, C. U., Gellert, M., & Bonner, W. M. (2000) · Current Biology · doi:10.1016/s0960-9822(00)00610-2 · Cited by 2,098
Self-citationKeeney, S., Giroux, C. N., & Kleckner, N. E. (1997) · Cell · doi:10.1016/s0092-8674(00)81876-0 · Cited by 1,824
Histone H2AX is phosphorylated in an ATR-dependent manner in response to replicational stress
Ward, I. M., & Chen, J. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.c100569200 · Cited by 1,150
Recombinational DNA double-strand breaks in mice precede synapsis
Mahadevaiah, S. K., Turner, J. M. A., Baudat, F., Rogakou, E. P., de Boer, P., Blanco‐Rodríguez, J., Jasin, M., Keeney, S., Bonner, W. M., & Burgoyne, P. S. (2001) · Nature Genetics · doi:10.1038/85830 · Cited by 852
Self-citationThe RAG Proteins and V(D)J Recombination: Complexes, Ends, and Transposition
Fugmann, S. D., Lee, A. I., Shockett, P. E., Villey, I. J., & Schatz, D. G. (2000) · Annual Review of Immunology · doi:10.1146/annurev.immunol.18.1.495 · Cited by 648
Haber, J. E. (2000) · Trends in Genetics · doi:10.1016/s0168-9525(00)02022-9 · Cited by 564
DNA double strand break repair and chromosomal translocation: Lessons from animal models
Ferguson, D. O., & Alt, F. W. (2001) · Oncogene · doi:10.1038/sj.onc.1204767 · Cited by 333
Response to RAG-Mediated V(D)J Cleavage by NBS1 and γ-H2AX
Chen, H. T., Bhandoola, A., Difilippantonio, M. J., Zhu, J., Brown, M. J., Tai, X., Rogakou, E. P., Brotz, T. M., Bonner, W. M., Ried, T., et al. (2000) · Science · doi:10.1126/science.290.5498.1962 · Cited by 326
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Evidence for a lack of DNA double-strand break repair in human cells exposed to very low x-ray doses
Rothkamm, K., & Löbrich, M. (2003) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0830918100 · Cited by 1,645
Bonner, W. M., Redon, C. E., Dickey, J. S., Nakamura, A. J., Sedelnikova, O. A., Solier, S., & Pommier, Y. (2008) · Nature reviews. Cancer · doi:10.1038/nrc2523 · Cited by 1,623
Self-citationγH2AX: a sensitive molecular marker of DNA damage and repair
Mah, L.-J., El-Osta, A., & Karagiannis, T. C. (2010) · Leukemia · doi:10.1038/leu.2010.6 · Cited by 1,557
-H2AX in recognition and signaling of DNA double-strand breaks in the context of chromatin
Kinner, A., Wu, W., Staudt, C., & Iliakis, G. E. (2008) · Nucleic Acids Research · doi:10.1093/nar/gkn550 · Cited by 1,303
Pathways of DNA Double-Strand Break Repair during the Mammalian Cell Cycle
Rothkamm, K., Krüger, I., Thompson, L. H., & Lübrich, M. (2003) · Molecular and Cellular Biology · doi:10.1128/mcb.23.16.5706-5715.2003 · Cited by 1,187
Metadata
- Authors
- Sedelnikova, O. A., Rogakou, E. P., Panyutin, I. G., & Bonner, W. M.
- Affiliations
- National Institutes of Health (US)
- National Cancer Institute (US)
- Corresponding author
- Sedelnikova, Olga A.
- Rogakou, Emmy P.
- Venue
- Radiation Research
- Publisher
- Radiation Research Society
- Published
- 2002-10
- Volume
- 158
- Issue
- 4
- Pages
- 486-492
- ISSN
- 0033-7587 (ISSN-L)
- 1938-5404
- OpenAlex
- W2106845421
- Cited by
- 611
- References
- 26
- Citations by year
- 2026: 4 · 2025: 8 · 2024: 10 · 2023: 17 · 2022: 14 · 2021: 22 · 2020: 17 · 2019: 22 · 2018: 17 · 2017: 24 · 2016: 34 · 2015: 24 · 2014: 26 · 2013: 28 · 2012: 35
- FWCI
- 7.18
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Radiation Research (publishedVersion)
- PubMed (publishedVersion)
- BioOne Complete (BioOne) (submittedVersion)
- 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
- Physical Sciences › Physics and Astronomy › Radiation › Radiation Therapy and Dosimetry
- Keywords
- DNA double-strand breaks, γH2AX, H2AX phosphorylation, DNA damage detection, iodine-125, HT1080 cells, radiation-induced DNA damage, nuclear foci
- MeSH terms
- Antibodies, DNA Damage, Histones, Humans, Idoxuridine, Iodine Radioisotopes, Tumor Cells, Cultured
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