Poly(ADP-Ribose)-Dependent Chromatin Remodeling in DNA Repair
| Autor: | Lebeaupin, Théo, Smith, Rebecca, Huet, Sébastien, Timinszky, Gyula |
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| Přispěvatelé: | Institut de Génétique et Développement de Rennes (IGDR), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Centre National de la Recherche Scientifique (CNRS)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES), Department of Physiological Chemistry, Butenandt Institute and Biomedical Center-Ludwig Maximilians University of Munich, Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Butenandt Institute and Biomedical Center-Ludwig-Maximilians University [Munich] (LMU) |
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
Poly Adenosine Diphosphate Ribose
[SDV.GEN]Life Sciences [q-bio]/Genetics DNA Repair Photo-activation Poly (ADP-Ribose) Polymerase-1 Chromatin Assembly and Disassembly PARP1 live-cell imaging Chromatin remodeling Poly ADP Ribosylation Animals Humans DNA Damage Response Poly(ADP-ribosyl)ation DNA Damage |
| Zdroj: | Methods in Molecular Biology Methods in Molecular Biology, Humana Press/Springer Imprint, 2017, 1608, pp.165-183. ⟨10.1007/978-1-4939-6993-7_12⟩ Methods in Molecular Biology, 2017, 1608, pp.165-183. ⟨10.1007/978-1-4939-6993-7_12⟩ |
| ISSN: | 1064-3745 1940-6029 |
| DOI: | 10.1007/978-1-4939-6993-7_12⟩ |
| Popis: | International audience; The tightly packed and dynamic structure of chromatin can undergo major reorganization in response to endogenous or exogenous stimuli, such as the regulation of transcription or the cell cycle, or following DNA damage. A fast and local chromatin decondensation is observed upon DNA damage induced by laser micro-irradiation. This decondensation is under the control of poly(ADP-ribosyl)ation (PARylation) by PARP1, one of the first proteins recruited at the DNA damage sites. This chapter provides a step-by-step guide to perform and analyze chromatin decondensation upon DNA damage induction. The protocol is based on fluorescence microscopy of live cells expressing a core histone tagged with a photoactivatable fluorophore. Laser micro-irradiation is used to simultaneously induce DNA damage and activate the fluorescence signal within the irradiated area. This photo-perturbation experiment can be easily implemented on any confocal laser-scanning microscope equipped with a photoperturbation module. The experimental framework can also be used to follow chromatin relaxation in parallel with the recruitment kinetics of a protein of interest at DNA lesions in cells co-expressing the tagged histones and a second protein of interest fused to a different fluorescent tag. |
| Databáze: | OpenAIRE |
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