Document Detail

Recombination by resolvase to analyse DNA communications by the SfiI restriction endonuclease.
MedLine Citation:
PMID:  8635479     Owner:  NLM     Status:  MEDLINE    
The SfiI endonuclease differs from other type II restriction enzymes by cleaving DNA concertedly at two copies of its recognition site, its optimal activity being with two sites on the same DNA molecule. The nature of this communication event between distant DNA sites was analysed on plasmids with recognition sites for SfiI interspersed with recombination sites for resolvase. These were converted by resolvase to catenanes carrying one SfiI site on each ring. The catenanes were cleaved by SfiI almost as readily as a single ring with two sites, in contrast to the slow reactions on DNA rings with one SfiI site. Interactions between SfiI sites on the same DNA therefore cannot follow the DNA contour and, instead, must stem from their physical proximity. In buffer lacking Mg2+, where SfiI is inactive while resolvase is active, the addition of SfiI to a plasmid with target sites for both proteins blocked recombination by resolvase, due to the restriction enzyme bridging its sites and thus isolating the sites for resolvase into separate loops. The extent of DNA looping by SfiI matched its extent of DNA cleavage in the presence of Mg2+.
M D Szczelkun; S E Halford
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The EMBO journal     Volume:  15     ISSN:  0261-4189     ISO Abbreviation:  EMBO J.     Publication Date:  1996 Mar 
Date Detail:
Created Date:  1996-07-11     Completed Date:  1996-07-11     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  8208664     Medline TA:  EMBO J     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  1460-9     Citation Subset:  IM    
Department of Biochemistry, University of Bristol, UK.
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MeSH Terms
Base Sequence
DNA / metabolism*
DNA Nucleotidyltransferases / metabolism*
Deoxyribonucleases, Type II Site-Specific / genetics,  metabolism*
Models, Genetic
Models, Molecular
Molecular Sequence Data
Nucleic Acid Conformation
Oligodeoxyribonucleotides / metabolism
Plasmids / metabolism
Recombinant Proteins / metabolism
Recombination, Genetic*
Grant Support
//Wellcome Trust
Reg. No./Substance:
0/Oligodeoxyribonucleotides; 0/Recombinant Proteins; 9007-49-2/DNA; EC 2.7.7.-/DNA Nucleotidyltransferases; EC 2.7.7.-/Transposases; EC 3.1.21.-/endodeoxyribonuclease SfiI; EC, Type II Site-Specific

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