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. 1985 Aug 26;13(16):5727–5746. doi: 10.1093/nar/13.16.5727

Investigation of restriction-modification enzymes from M. varians RFL19 with a new type of specificity toward modification of substrate.

V Butkus, S Klimasauskas, D Kersulyte, D Vaitkevicius, A Lebionka, A Janulaitis
PMCID: PMC321908  PMID: 2994011

Abstract

The characterization of MvaI restriction-modification enzymes, isolated from Micrococcus varians RFL19, is reported. Both enzymes recognize the 5'CC decreases (A/T)GG nucleotide sequence. The endonuclease cleaves the sequence at the position indicated by the arrow, whereas the methylase modifies the internal cytosine, yielding N4-methylcytosine. This type of modification protects the substrate from R.MvaI cleavage. 5-Methylcytosine in the same position of the recognition sequence does not protect the substrate from R.MvaI cleavage. R.MvaI proved to be the first example of a restriction endonuclease differentiating the position of the methyl group in the heterocyclic ring of cytosine, located in the same site of the recognition sequence. M.MvaI modifies DNA dcm+ in vitro yielding N4,5-dimethylcytosine. N4-methylcytosine cannot be differentiated from cytosine using the Maxam-Gilbert DNA sequencing procedure.

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