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J Biol Chem, Vol. 274, Issue 51, 36379-36386, December 17, 1999
From the Department of Biochemistry, School of Medical Sciences,
University of Bristol, University Walk,
Bristol BS8 1TD, United Kingdom
Type II restriction endonucleases usually
recognize 4-6-base pair (bp) sites on DNA and cleave each site in a
separate reaction. A few type II endonucleases have 8-bp recognition
sites, but these seem unsuited for restriction, since their sites are
rare on most DNA. Moreover, only one endonuclease that recognizes a
target containing 8 bp has been examined to date, and this enzyme,
SfiI, needs two copies of this site for its DNA cleavage
reaction. In this study, several endonucleases with 8-bp sites were
tested on plasmids that have either one or two copies of the relevant sequence to determine if they also need two sites. SgfI,
SrfI, FseI, PacI, PmeI,
Sse8781I, and SdaI all acted through equal and independent reactions at each site. AscI cleaved the DNA
with one site at the same rate as that with two sites but acted
processively on the latter. In contrast, SgrAI showed a
marked preference for the plasmid with two sites and cleaved both sites
on this DNA in a concerted manner, like SfiI. Endonucleases
that require two copies of an 8-bp sequence may be widespread in
nature, where, despite this seemingly inappropriate requirement, they
may function in DNA restriction.
Reactions of Type II Restriction Endonucleases with 8-Base
Pair Recognition Sites*
*
This work was supported by grants from the Wellcome Trust
and the Biotechnology and Biological Sciences Research Council.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of Biochemistry,
School of Medical Sciences, University of Bristol, University Walk,
Bristol BS8 1TD, UK. Tel.: 44-117-928-7429; Fax: 44-117-928-8274; E-mail: s.halford@bris.ac.uk.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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