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

• Published in: Methods 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
• Language: English
• 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
• DOI: 10.1007/978-1-4939-2474-5_19

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.