Cited by 2,422DNA Repair MechanismsEffects of Radiation ExposureGenomics and Chromatin Dynamics

Megabase Chromatin Domains Involved in DNA Double-Strand Breaks in Vivo

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Rogakou, Emmy P., Boon, Chye, Redon, Christophe, Bonner, William M.·1999·The Journal of Cell Biology

Abstract

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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  1. Celis J.E. Cell Biology. Vol. 2. 2nd ed Academic Press 1998 533 San Diego CApp
  2. Rathmell W.K. Chu G. Mechanisms for DNA double-strand break repair in eucaryotes Nickoloff J.A. Hoekstra M.F. DNA Damage and Repair. Vol. 2 1998 299 316 Humana Press Totowa, NJ

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Authors
Rogakou, E. P., Boon, C., Redon, C., & Bonner, W. M.
Affiliations
National Institutes of Health (US)
Venue
The Journal of Cell Biology
Published
1999-09-06
Volume
146
Issue
5
Pages
905-916
ISSN
  • 0021-9525 (ISSN-L)
  • 1540-8140
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
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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