Document Detail


DNA binding and recognition by the IIs restriction endonuclease MboII.
MedLine Citation:
PMID:  11606594     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The type IIs restriction endonuclease MboII recognizes nonsymmetrical GAAGA sites, cutting 8 (top strand) and 7 (bottom strand) bases to the right. Gel retardation showed that MboII bound specifically to GAAGA sequences, producing two distinct complexes each containing one MboII and one DNA molecule. Interference analysis indicated that the initial species formed, named complex 1, comprised an interaction between the enzyme and the GAAGA target. Complex 2 involved interaction of the protein with both the GAAGA and the cutting sites. Only in the presence of divalent metal ions such as Ca(2+) is the conversion of complex 1 to 2 rapid. Additionally, a very retarded complex was seen with Ca(2+), possibly a (MboII)(2)-(DNA)(2) complex. Plasmids containing a single GAAGA site were hydrolyzed slowly by MboII. Plasmids containing two sites were cut far more rapidly, suggesting that the enzyme requires two recognition sites in the same DNA molecule for efficient hydrolysis. MboII appears to have a mechanism similar to the best characterized type IIs enzyme, FokI. Both enzymes initially bind DNA as monomers, followed by dimerization to give an (enzyme)(2)-(DNA)(2) complex. Dimerization is efficient only when the two target sites are located in the same DNA molecule and requires divalent metal ions.
Authors:
Meera Soundararajan; Zhiyuh Chang; Richard D Morgan; Pauline Heslop; Bernard A Connolly
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2001-10-17
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  277     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2002 Jan 
Date Detail:
Created Date:  2002-01-07     Completed Date:  2002-02-07     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  887-95     Citation Subset:  IM    
Affiliation:
Department of Biochemistry and Molecular Genetics, The University of Newcastle, Newcastle upon Tyne, NE2 4HH, United Kingdom.
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MeSH Terms
Descriptor/Qualifier:
Calcium / metabolism
DNA / metabolism*
Deoxyribonucleases, Type II Site-Specific / isolation & purification,  metabolism*
Dimerization
Magnesium / metabolism
Oligonucleotides / metabolism*
Plasmids
Chemical
Reg. No./Substance:
0/Oligonucleotides; 7439-95-4/Magnesium; 7440-70-2/Calcium; 9007-49-2/DNA; EC 3.1.21.-/endodeoxyribonuclease MboII; EC 3.1.21.4/Deoxyribonucleases, Type II Site-Specific

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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