ChIP-chip profiling of human gammaH2AX spreading around DNA double strand breaks
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ABSTRACT: Chromatin acts as a key regulator of DNA related processes such as DNA damage repair. While ChIP-chip is a powerful technique to provide high-resolution maps of protein-genome interactions, its use to study DNA Double Strand Break (DSB) repair has been hindered by the limitations of the available damage induction methods. We have developed a human cell line that permits induction of multiple DSBs randomly distributed and unambiguously positioned within the genome. Using this system, we have generated the first genome-wide mapping of gammaH2AX around DSBs. We found that all DSBs trigger large gammaH2AX domains, which extend from the DSB in a bidirectional, discontinuous and not necessarily symmetrical manner. Strikingly we uncovered that, within domains, gammaH2AX distribution is highly influenced by gene transcription since parallel mapping of RNA Polymerase II and strand specific expression revealed that ?H2AX does not propagate on active genes. In addition, we demonstrate that transcription is accurately maintained within gammaH2AX domains, indicating that mechanisms may exist to protect genes transcription from gammaH2AX spreading and from the chromatin rearrangements induced by DSBs.
ORGANISM(S): Homo sapiens
SUBMITTER: gaelle legube
PROVIDER: E-MEXP-1769 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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