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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
10
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pubmed:dateCreated |
1988-5-2
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pubmed:abstractText |
The DNA methyltransferase M-BsuE that recognizes the sequence 5'-CGCG-3' has been isolated from Bacillus subtilis strain ISE15. A 1600-fold purification of M-BsuE was achieved by column chromatography on phosphocellulose, heparin-Sepharose, and DEAE-Sepharose. DNA methyltransferase activity was monitored in the column eluants radiochemically by the transfer of tritiated methyl groups from radiolabeled S-adenosylmethionine to poly(dGdC)-poly(dGdC) DNA, a sensitive and specific substrate for M-BsuE activity. The DNA sequence specificity of this methyltransferase activity was confirmed enzymatically by demonstrating that M-BsuE-methylated DNA was selectively protected from cleavage by the restriction enzyme isoschizomers, ThaI and FnuDII. Purified M-BsuE has an apparent molecular size of 41,000-43,000 as determined by gel filtration and migrates as a 41-kDa protein in a sodium dodecyl sulfate-polyacrylamide gel. DNA methylation by M-BsuE is dependent upon the presence of S-adenosylmethionine and 2-mercaptoethanol. M-BsuE methyltransferase activity is optimal at 37 degrees C in the presence of 50 mM Tris-HCl, pH 7.8, 25 mM KCl, 6 microM S-adenosylmethionine, 5 mM 2-mercaptoethanol, and 10 mM EDTA. M-BsuE methylates the external cytidine in its recognition sequence in both linear and supercoiled DNA. A unique property of M-BsuE is its ability to methylate 5'-CGCG-3' in Z-DNA.
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pubmed:grant | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Apr
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pubmed:issn |
0021-9258
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
5
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pubmed:volume |
263
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
4832-6
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pubmed:dateRevised |
2009-11-19
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pubmed:meshHeading |
pubmed-meshheading:3127393-Bacillus subtilis,
pubmed-meshheading:3127393-Base Sequence,
pubmed-meshheading:3127393-Chromatography, Affinity,
pubmed-meshheading:3127393-Chromatography, Ion Exchange,
pubmed-meshheading:3127393-DNA-Cytosine Methylases,
pubmed-meshheading:3127393-Kinetics,
pubmed-meshheading:3127393-Methylation,
pubmed-meshheading:3127393-Methyltransferases,
pubmed-meshheading:3127393-Osmolar Concentration,
pubmed-meshheading:3127393-Substrate Specificity
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pubmed:year |
1988
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pubmed:articleTitle |
Isolation and characterization of BsuE methyltransferase, a CGCG specific DNA methyltransferase from Bacillus subtilis.
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pubmed:affiliation |
Department of Pharmacology and Toxicology, West Virginia University, Morgantown 26506.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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