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Modification methylase M.Sau3239I from Streptomyces aureofaciens 3239
E Zelinková1, M Paulícek, J Zelinka
1Institute of Molecular Biology, Slovak Academy of Sciences, Bratislava, Czechoslovakia.
Abstract:
By chromatography on phosphocellulose and Heparin-Sepharose the modification methylase M.Sau3239I was detected and partly purified from cells of Streptomyces aureofaciens 3239. Methylation by this enzyme protects DNA from cleavage by the restriction endonuclease R.Sau3239I. The enzyme catalyzes methylation of adenine to N-6-methyladenine in the 5'-CTCGmAG-3' recognition sequence.
Insights
Researchers purified the modification methylase M.Sau3239I from Streptomyces aureofaciens. This enzyme methylates adenine in DNA, protecting it from restriction endonuclease R.Sau3239I cleavage.
Area of Science:
- Molecular Biology
- Enzymology
- Microbial Genetics
Background:
- Restriction-modification systems are crucial for bacterial defense and genome regulation.
- DNA methylation plays a key role in protecting host DNA from restriction enzymes.
Purpose of the Study:
- To detect and partially purify the modification methylase M.Sau3239I from Streptomyces aureofaciens.
- To characterize the enzymatic activity and recognition sequence of M.Sau3239I.
Main Methods:
- Chromatography techniques, including phosphocellulose and Heparin-Sepharose, were employed for enzyme purification.
- Enzyme activity was assessed by its ability to methylate DNA and confer protection against restriction digestion.
Main Results:
- The modification methylase M.Sau3239I was successfully detected and partially purified.
- The enzyme was identified as catalyzing the methylation of adenine to N-6-methyladenine.
- The specific recognition sequence for M.Sau3239I was determined to be 5'-CTCGmAG-3'.
Conclusions:
- M.Sau3239I is a DNA modification methylase from Streptomyces aureofaciens.
- The enzyme's activity involves methylation at adenine within the 5'-CTCGmAG-3' sequence.
- This methylation event confers protection against the cognate restriction endonuclease R.Sau3239I.