Dynamics of a DNA mismatch site held in confinement discriminate epigenetic modifications of cytosine

Johnson, Robert P., Fleming, Aaron M., Perera, Rukshan T. , Burrows, Cynthia J. and White, Henry S. (2017) Dynamics of a DNA mismatch site held in confinement discriminate epigenetic modifications of cytosine. Journal of the American Chemical Society, 139 (7). pp. 2750-2756. ISSN 0002-7863

Full content URL: http://doi.org/10.1021/jacs.6b12284

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Dynamics of a DNA mismatch site held in confinement discriminate epigenetic modifications of cytosine
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Abstract

The identification and discrimination of four epigenetic modifications to cytosine in the proposed active demethylation cycle is demonstrated at the single-molecule level, without the need for chemical pretreatment or labeling. The wild-type protein nanopore α-hemolysin is used to capture individual DNA duplexes containing a single cytosine–cytosine mismatch. The mismatch is held at the latch constriction of α-hemolysin, which is used to monitor the kinetics of base-flipping at the mismatch site. Base-flipping and the subsequent interactions between the DNA and the protein are dramatically altered when one of the cytosine bases is replaced with methyl-, hydroxymethyl-, formyl-, or carboxylcytosine. As well as providing a route to single-molecule analysis of important epigenetic markers in DNA, our results provide important insights into how the introduction of biologically relevant, but poorly understood, modifications to cytosine affect the local conformational dynamics of a DNA duplex in a confined environment.

Keywords:Hemolysin, Epigenetics, DNA
Subjects:F Physical Sciences > F170 Physical Chemistry
Divisions:College of Science > School of Chemistry
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ID Code:27423
Deposited On:27 Apr 2017 19:26

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