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PMID: 16520385 Published · ppublish English Journal Article Research Support, N.I.H., Intramural Research Support, Non-U.S. Gov't

Changes in chromatin structure and mobility in living cells at sites of DNA double-strand breaks.

The Journal of cell biology ·Vol. 172 ·No. 6 ·2006-03-13 ·Pages 823-34

Kruhlak MJ, Celeste A, Dellaire G, Fernandez-Capetillo O, Müller WG, McNally JG, Bazett-Jones DP, Nussenzweig A

Abstract

The repair of DNA double-strand breaks (DSBs) is facilitated by the phosphorylation of H2AX, which organizes DNA damage signaling and chromatin remodeling complexes in the vicinity of the lesion. The disruption of DNA integrity induces an alteration of chromatin architecture that has been proposed to activate the DNA damage transducing kinase ataxia telangiectasia mutated. However, little is known about the physical properties of damaged chromatin. In this study, we use a photoactivatable version of GFP-tagged histone H2B to examine the mobility and structure of chromatin containing DSBs in living cells. We find that chromatin containing DSBs exhibits limited mobility but undergoes an energy-dependent local expansion immediately after DNA damage. The localized expansion observed in real time corresponds to a 30-40% reduction in the density of chromatin fibers in the vicinity of DSBs, as measured by energy-filtering transmission electron microscopy. The observed opening of chromatin occurs independently of H2AX and ATM. We propose that localized adenosine triphosphate-dependent decondensation of chromatin at DSBs establishes an accessible subnuclear environment that facilitates DNA damage signaling and repair.

MeSH Terms
Adenosine Triphosphate/metabolism Animals Cells, Cultured Chromatin/chemistry,genetics,ultrastructure Chromosome Positioning/genetics DNA/genetics,ultrastructure DNA Damage/genetics DNA Repair/genetics Energy Metabolism/genetics Female Fibroblasts Green Fluorescent Proteins HeLa Cells Histones/genetics,metabolism Humans Male Mice Mice, Knockout Microscopy, Electron, Transmission Signal Transduction/genetics
Chemicals
Chromatin Histones Green Fluorescent Proteins Adenosine Triphosphate DNA
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Kruhlak Michael J
Experimental Immunology Branch and 2Laboratory for Receptor Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. kruhlakm@mail.nih.gov
Celeste Arkady
Dellaire Graham
Fernandez-Capetillo Oscar
Müller Waltraud G
McNally James G
Bazett-Jones David P
Nussenzweig André
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200510015
Email
kruhlakm@mail.nih.gov
Published
2006-03-13
Epub
2006-00-06
Pages
823-34
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2063727
Subset
IM
Grants
Intramural NIH HHS · United States
External Links
PubMed source DOI: 10.1083/jcb.200510015 PMC Full Text View journal details
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