Megabase Chromatin Domains Involved in DNA Double-Strand Breaks in Vivo
Open accessAbstract
The loss of chromosomal integrity from DNA double-strand breaks introduced into mammalian cells by ionizing radiation results in the specific phosphorylation of histone H2AX on serine residue 139, yielding a specific modified form named γ-H2AX. An antibody prepared to the unique region of human γ-H2AX shows that H2AX homologues are phosphorylated not only in irradiated mammalian cells but also in irradiated cells from other species, including Xenopus laevis, Drosophila melanogaster, and Saccharomyces cerevisiae. The antibody reveals that γ-H2AX appears as discrete nuclear foci within 1 min after exposure of cells to ionizing radiation. The numbers of these foci are comparable to the numbers of induced DNA double-strand breaks. When DNA double-strand breaks are introduced into specific partial nuclear volumes of cells by means of a pulsed microbeam laser, γ-H2AX foci form at these sites. In mitotic cells from cultures exposed to nonlethal amounts of ionizing radiation, γ-H2AX foci form band-like structures on chromosome arms and on the end of broken arms. These results offer direct visual confirmation that γ-H2AX forms en masse at chromosomal sites of DNA double-strand breaks. The results further suggest the possible existence of units of higher order chromatin structure involved in monitoring DNA integrity.
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- Authors
- Rogakou, E. P., Boon, C., Redon, C., & Bonner, W. M.
- Affiliations
- National Institutes of Health (US)
- ORCID
- Redon, Christophe
- Venue
- The Journal of Cell Biology
- Publisher
- Rockefeller University Press
- Published
- 1999-09-06
- Volume
- 146
- Issue
- 5
- Pages
- 905-916
- ISSN
- 0021-9525 (ISSN-L)
- 1540-8140
- PMC
- PMC2169482
- OpenAlex
- W2084969545
- Cited by
- 2,422
- References
- 30
- Citations by year
- 2026: 26 · 2025: 44 · 2024: 53 · 2023: 55 · 2022: 69 · 2021: 96 · 2020: 94 · 2019: 87 · 2018: 74 · 2017: 94 · 2016: 107 · 2015: 110 · 2014: 116 · 2013: 109 · 2012: 121
- FWCI
- 9.88
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- The Journal of Cell Biology (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-sa)
- PubMed Central (publishedVersion, cc-by-nc-sa)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- National Cancer Institute
- National Institute of Child Health and Human Development
- 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 › Molecular Biology › Genomics and Chromatin Dynamics
- Keywords
- DNA double-strand breaks, γH2AX, H2AX phosphorylation, ionizing radiation, chromatin domains, nuclear foci, chromosome integrity, mitosis, chromatin structure, DNA damage response
- MeSH terms
- Animals, Cell Line, Chromatin, Chromosomes, DNA, DNA Damage, Fibroblasts, Histones, Humans, Interphase, Lasers, Mitosis, Phosphorylation, Phosphoserine, Radiation, Ionizing, Time Factors, Blotting, Western, Microscopy, Confocal, Muntjacs
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