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A Mutation in the Mod Subunit of EcoP15I Restriction Enzyme Converts the DNA Methyltransferase to a Site-specific Endonuclease

Bist, Pradeep and Madhusoodanan, Urulangodi Kunhiraman and Rao, Desirazu N (2007) A Mutation in the Mod Subunit of EcoP15I Restriction Enzyme Converts the DNA Methyltransferase to a Site-specific Endonuclease. In: Journal of Biological Chemistry, 282 (6). pp. 3520-3530.

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Abstract

A closer inspection of the amino acid sequence of EcoP15I DNA methyltransferase revealed a region of similarity to the $PDX_n(D/E)XK$ catalytic site of type II restriction endonucleases, except for methionine in EcoP15I DNA methyltransferase instead of proline. Substitution of methionine at position 357 by proline converts EcoP15I DNA methyltransferase to a site-specific endonuclease. EcoP15I-M357P DNA methyltransferase specifically binds to the recognition sequence 5'-CAGCAG-3' and cleaves DNA asymmetrically EcoP151-M357P·DNA methyltransferase specifically binds to the recognition sequence 5'-CAGCAG-3' and cleaves DNA asymmetrically, $5'-CAGCAG(N)_{10}-3'$, as indicated by the arrows, in presence of magnesium ions.

Item Type: Journal Article
Additional Information: Copyright of this article belongs to the American Society for Biochemistry and Molecular Biology.
Department/Centre: Division of Biological Sciences > Biochemistry
Date Deposited: 09 Apr 2007
Last Modified: 27 Aug 2008 12:45
URI: http://eprints.iisc.ernet.in/id/eprint/10535

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