Imaging local deposition of newly synthesized histones in UVC-damaged chromatin

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...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 1288; p. 337
Main Authors Adam, Salomé, Dabin, Juliette, Bai, Siau-Kun, Polo, Sophie E
Format Journal Article
LanguageEnglish
Published United States 01.01.2015
Subjects
Cell Line
Chromatin - genetics
Chromatin - metabolism
DNA Damage - radiation effects
Histones - biosynthesis
Humans
Microscopy, Fluorescence
S-Nitroso-N-Acetylpenicillamine - chemistry
Staining and Labeling - methods
Ultraviolet Rays - adverse effects
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Summary: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.
ISSN:1940-6029
DOI:10.1007/978-1-4939-2474-5_19