Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.1.1.37 (DNA methyltransferase)
 4,983 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We present a novel strategy with synthetic hemimethylated DNA substrates containing uracil for thymine and inosine for guanosine replacements and EcoRI DNA methyltransferase to characterize the importance of major groove hydrophobic groups to the sequence-specific modification of DNA. The bacterial Mtase uses S-adenosyl-L-methionine to methylate the double-stranded DNA site 5'GAATTC3' at the N6 position of the central adenosine of each strand. Uracil substitution in either strand at the outer thymine (5'GAATUC3') causes 2.2- and 1.7-fold improvements in specificity (kcat/KmDNA). The fact that the specificity constant for the substrate containing uracil in both strands is identical to the value expected for noninteracting substitutions suggests that no significant methyltransferase-DNA interactions are altered beyond the site of either substitution. Similar analysis of the internal thymine (5'GAAUTC3') also shows these methyl groups to make a negative contribution to specificity, although the observed nonadditivity with the doubly modified substrate clearly shows methyltransferase-DNA interactions beyond the site of substitution to be affected in this case. To further probe the effect of analogue incorporation on methyltransferase-DNA interactions beyond the site of substitution, the relatively "silent" and additive uracil changes (5'GAATUC3') were combined with inosine for guanosine substitutions (e.g., 5'IAATTC3') known to have significant negative effects on specificity. In contrast to the additivity observed with the outer thymines, these studies show significant changes in methyltransferase-DNA interactions caused by the removal of the thymine methyls. Our results implicate a complex and flexible methyltransferase-DNA interface in which subtle structural changes in the substrate are transmitted over the entire canonical site.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:EcoRI DNA methyltransferase-DNA interactions. 153 35

In Bacillus subtilis, 5-azacytidine, an analog of cytidine, causes a time- and dose-dependent growth inhibition. Methyl donors are unable to overcome azacytidine-induced inhibition while pyrimidine nucleosides, except orotidine, can revert this inhibition totally. On the other hand, pyrimidine bases, except uracil, are unable to restore growth in azacytidine-treated cells. Uracil, at a high concentration, can revert growth inhibition only inefficiently. However, a considerable relief of growth inhibition by uracil occurs in the presence of a ribose donor. In azacytidine-treated B. subtilis cells methylation of bases in DNA is not affected either quantitatively or qualitatively and DNA methyltransferase activity remains unaltered as compared to the untreated cells, apparently due to the absence of azacytidine incorporation into the DNA. The inability of B. subtilis cytidine kinase to phosphorylate azacytidine is the probable reason for this non-incorporation. Analysis of the enzymes of de novo pyrimidine biosynthesis has shown that orotidine monophosphate pyrophosphorylase is specifically repressed by azacytidine treatment.
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PMID:Effect of 5-azacytidine on DNA methylation and on the enzymes of de novo pyrimidine biosynthesis in Bacillus subtilis Marburg strain. 620 29