期刊
DNA REPAIR
卷 20, 期 -, 页码 49-57出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.dnarep.2014.02.018
关键词
Single-molecule; Magnetic trapping; Transcription; Repair; Helicase
资金
- Centre National de la Recherche Scientifique
- University of Paris 7 - Paris Diderot
- EURYI award
- French Ministry of Research
- Association pour la Recherche contre le Cancer
- China Science Council
DNA repair is often a complex, multi-component, multi-step process; this makes detailed kinetic analysis of the different steps of repair a challenging task using standard biochemical methods. At the same time, single-molecule methods are well-suited for extracting kinetic information despite time-averaging due to diffusion of biochemical components and stochasticity of chemical reaction steps. Here we discuss recent experiments using DNA nanomanipulation in a magnetic trap to study the initiation of transcription-coupled repair in a model bacterial system comprising the canonical Escherichia coli RNA polymerase and the Mfd translocase which specifically binds to it. These experiments provide kinetic insight into the reaction process, helping to explain how Mfd discriminates between transcribing RNAP and stalled RNAP. They also identify a reliably long-lived intermediate containing Mfd translocase and, potentially, RNA polymerase. This intermediate presumably serves as a platform for assembly of downstream repair components UvrAB(C). (C) 2014 Elsevier B.V. All rights reserved.
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