Abstract
DNA damage not only jeopardizes genome integrity but also challenges the well-organized association of DNA with histone proteins into chromatin, which is key for regulating gene expression and cell functions. The extent to which the original chromatin structure is altered after repair of DNA lesions is thus a critical issue. Dissecting histone dynamics at sites of DNA damage has provided mechanistic insights into chromatin plasticity in response to genotoxic stress. Here, we present an experimental protocol for visualizing the deposition of newly synthesized histone H3 variants at sites of UVC damage in human cells that couples SNAP-tag based labeling of new histones with local UVC irradiation of cells through micropore filters.
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Acknowledgements
We thank Dominique Ray-Gallet for critical reading of the manuscript. Research in S.E.P. group is supported by the European Research Council (ERC starting grant ERC-2013-StG-336427 “EpIn”), the French National Research Agency (ANR-12-JSV6-0002-01), the “Who am I?” laboratory of excellence (ANR-11-LABX-0071) funded by the French Government through its “Investments for the Future” program (ANR-11-IDEX-0005-01), EDF Radiobiology program RB 2014-01 and the Foundation ARC. S.A. is recipient of a PhD fellowship from University Pierre and Marie Curie and La Ligue contre le Cancer.
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Adam, S., Dabin, J., Bai, SK., Polo, S.E. (2015). Imaging Local Deposition of Newly Synthesized Histones in UVC-Damaged Chromatin. In: Chellappan, S. (eds) Chromatin Protocols. Methods in Molecular Biology, vol 1288. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2474-5_19
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DOI: https://doi.org/10.1007/978-1-4939-2474-5_19
Publisher Name: Humana Press, New York, NY
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