M.BstF5I-2 and M.BstF5I-4 DNA methyltransferases from BstF5I restriction-modification system of Bacillus stearothermophilus F5.

V A Chernukhin, L N Golikova, D A Gonchar, M A Abdurashitov, Yu G Kashirina, N A Netesova, S Kh Degtyarev

Institute of Molecular Biology and Biophysics, Siberian Division, Russian Academy of Medical Sciences, Novosibirsk 630117, Russia.

Biochemistry. Biokhimii͡a 2003 Sep

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The BstF5I restriction-modification system from Bacillus stearothermophilus F5 includes four site-specific DNA methyltransferases, thus differing from all known restriction-modification systems. Here we demonstrated for the first time that one bacterial cell can possess two pairs of methylases with identical substrate specificities (methylases BstF5I-1 and BstF5I-3 recognize GGATG, whereas methylases BstF5I-2 and BstF5I-4 recognize CATCC) that modify adenine residues on both DNA strands. Different chromatographic methods provide homogenous preparations of methylases BstF5I-2 and BstF5I-4. We estimated the principal kinetic parameters of the reaction of transfer of methyl group from the donor S-adenosyl-L-methionine to the recognition site 5;-CATCC-3; catalyzed by BstF5I-2 and BstF5I-4 DNA [N6-adenine]-methyltransferases from the BstF5I restriction-modification system.

Citation

V A Chernukhin, L N Golikova, D A Gonchar, M A Abdurashitov, Yu G Kashirina, N A Netesova, S Kh Degtyarev. M.BstF5I-2 and M.BstF5I-4 DNA methyltransferases from BstF5I restriction-modification system of Bacillus stearothermophilus F5. Biochemistry. Biokhimii͡a. 2003 Sep;68(9):967-75