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Single-Molecule Methods for Nucleotide Excision Repair: Building a System to Watch Repair in Real Time

Kong, Muwen and Beckwitt, Emily C and Springall, Luke and Kad, Neil M and Van Houten, Bennett (2017) Single-Molecule Methods for Nucleotide Excision Repair: Building a System to Watch Repair in Real Time. In: Methods in Enzymology. Elsevier, pp. 213-257. ISBN 978-0-12-811469-8. (doi:10.1016/bs.mie.2017.03.027) (Access to this publication is currently restricted. You may be able to access a copy if URLs are provided) (KAR id:65159)

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https://doi.org/10.1016/bs.mie.2017.03.027

Abstract

Single-molecule approaches to solving biophysical problems are powerful tools that allow static and dynamic real-time observations of specific molecular interactions of interest in the absence of ensemble-averaging effects. Here, we provide detailed protocols for building an experimental system that employs atomic force microscopy and a single-molecule DNA tightrope assay based on oblique angle illumination fluorescence microscopy. Together with approaches for engineering site-specific lesions into DNA substrates, these complementary biophysical techniques are well suited for investigating protein–DNA interactions that involve target-specific DNA-binding proteins, such as those engaged in a variety of DNA repair pathways. In this chapter, we demonstrate the utility of the platform by applying these techniques in the studies of proteins participating in nucleotide excision repair.

Item Type: Book section
DOI/Identification number: 10.1016/bs.mie.2017.03.027
Subjects: Q Science
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: N. Kad
Date Deposited: 10 Dec 2017 11:24 UTC
Last Modified: 29 May 2019 20:01 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/65159 (The current URI for this page, for reference purposes)
Kad, Neil M: https://orcid.org/0000-0002-3491-8595
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