Binding of DNA Methyltransferase M.Ecl18 to Operator–Promoter Region Decreases Its Methylating Activity

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D. V. Nikitin^*, M. L. Mokrishcheva, and A. S. Solonin

Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, pr. Nauki 5, 142290 Pushchino, Moscow Region, Russia; fax: (495) 956-3370; E-mail: dvnikitin@rambler.ru

^* To whom correspondence should be addressed.

Received November 17, 2011; Revision received December 2, 2011

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The type II bifunctional DNA methyltransferase (MTase) Ecl18 that is able to control transcription of its own gene was studied kinetically. Based on initial velocity dependences from S-adenosyl-L-methionine (AdoMet) and target DNA and substrate preincubation assays, it is proposed that the enzyme apparently works by a rapid equilibrium ordered bi-bi mechanism with DNA binding first. By measuring the enzyme activity depending on DNA and AdoMet at different fixed concentrations of the operator sequence oligonucleotide, it was found that its binding has noncompetitive inhibitory effect on Ecl18 MTase activity.

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KEY WORDS: type II DNA methyltransferase, enzyme kinetics, noncompetitive inhibition

DOI: 10.1134/S0006297912030108

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