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Sequence-specific double-strand cleavage of DNA by penta-N-methylpyrrolecarboxamide-EDTA·Fe(II)

Schultz, Peter G. and Dervan, Peter B. (1983) Sequence-specific double-strand cleavage of DNA by penta-N-methylpyrrolecarboxamide-EDTA·Fe(II). Proceedings of the National Academy of Sciences of the United States of America, 80 (22). pp. 6834-6837. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:SCHpnas83

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Abstract

In the presence of O2 and 5 mM dithiothreitol, penta-N-methylpyrrolecarboxamide-EDTA·Fe(II) [P5E·Fe(II)] at 0.5 µ M cleaves pBR322 plasmid DNA (50 µ M in base pairs) on opposite strands to afford discrete DNA fragments as analyzed by agarose gel electrophoresis. High-resolution denaturing gel electrophoresis of a 32P-end-labeled 517-base-pair restriction fragment containing a major cleavage site reveals that P5E·Fe(II) cleaves 3-5 base pairs contiguous to a 6-base-pair sequence, 5'-T-T-T-T-T-A-3' (4,323-4,328 base pairs). The major binding orientation of the pentapeptide occurs with the amino terminus at the adenine side of this sequence. In the presence of 5 mM dithiothreitol, 0.01 µ M P5E·Fe(II) converts form I pBR322 DNA at 0.22 µ M plasmid (1.0 mM in base pairs) to 40% form II, indicating the cleavage reaction is catalytic, turning over a minimum of nine times. This synthetic molecule achieves double-strand cleavage of DNA (pH 7.9, 25 degrees C) at the 6-base-pair recognition level and may provide an approach to the design of "artificial restriction enzymes."


Item Type:Article
Additional Information:Copyright © 1983 by the National Academy of Sciences. Communicated by John J. Hopfield, August 8, 1983. We are grateful to the National Institutes of Health for grant support (GM-27681) and to IBM for a fellowship to P.G.S. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Subject Keywords:minor groove binding; netropsin; distamycin; restriction enzymes
Record Number:CaltechAUTHORS:SCHpnas83
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:SCHpnas83
Alternative URL:http://www.pnas.org/cgi/content/abstract/80/22/6834
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:1559
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:30 Jan 2006
Last Modified:14 Nov 2014 19:19

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