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Nucleic Acids Research Advance Access published online on April 28, 2009
Nucleic Acids Research, doi:10.1093/nar/gkp267
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© 2009 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Nucleic Acid Enzymes

S-Adenosyl homocysteine and DNA ends stimulate promiscuous nuclease activities in the Type III restriction endonuclease EcoPI

Luke J. Peakman and Mark D. Szczelkun*

DNA–Protein Interactions Unit, Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, UK

*To whom correspondence should be addressed. Tel: +44 117 331 2158; Fax: +44 117 331 2168; Email: mark.szczelkun{at}bristol.ac.uk

Received March 6, 2009. Revised April 9, 2009. Accepted April 9, 2009.

In the absence of the methyl donor S-adenosyl methionine and under certain permissive reaction conditions, EcoPI shows non-specific endonuclease activity. We show here that the cofactor analogue S-adenosyl homocysteine promotes this promiscuous DNA cleavage. Additionally, an extensive exonuclease-like processing of the DNA is also observed that can even result in digestion of non-specific DNA in trans. We suggest a model for how DNA communication events initiating from non-specific sites, and in particular free DNA ends, could produce the observed cleavage patterns.

Present address: Luke J. Peakman, South Devon College, Vantage Point, Long Road, Paignton, Devon TQ4 7EJ, UK


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