Nucleic Acids Research, 2000, Vol. 28, No. 20 3950-3961
© 2000 Oxford University Press
Structure of RsrI methyltransferase, a member of the N6-adenine ß class of DNA methyltransferases
Department of Pharmacology, University of Colorado Health Sciences Center, 4200 East Ninth Avenue, Denver, CO 80262, USA, 1Department of Biochemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, Urbana, IL 61801, USA and 2Structural Biology Center, Biosciences Division, Argonne National Laboratories, 9700 South Cass Avenue, Argonne, IL 60439-4833, USA
DNA methylation is important in cellular, developmental and disease processes, as well as in bacterial restriction–modification systems. Methylation of DNA at the amino groups of cytosine and adenine is a common mode of protection against restriction endonucleases afforded by the bacterial methyltransferases. The first structure of an N6-adenine methyltransferase belonging to the ß class of bacterial methyltransferases is described here. The structure of M·RsrI from Rhodobacter sphaeroides, which methylates the second adenine of the GAATTC sequence, was determined to 1.75 Å resolution using X-ray crystallography. Like other methyltransferases, the enzyme contains the methylase fold and has well-defined substrate binding pockets. The catalytic core most closely resembles the PvuII methyltransferase, a cytosine amino methyltransferase of the same ß group. The larger nucleotide binding pocket observed in M·RsrI is expected because it methylates adenine. However, the most striking difference between the RsrI methyltransferase and the other bacterial enzymes is the structure of the putative DNA target recognition domain, which is formed in part by two helices on an extended arm of the protein on the face of the enzyme opposite the active site. This observation suggests that a dramatic conformational change or oligomerization may take place during DNA binding and methylation.
* To whom correspondence should be addressed. Tel: +1 303 315 0427; Fax: +1 303 315 7097; Email: mair.churchill@uchsc.edu Present addresses: Sandra S. Szegedi, Department of Chemistry and Biochemistry, University of California at Santa Barbara, Santa Barbara, CA 93106, USA Martin A. Walsh, Istituto di Richerche di Biologia Molecolare, Via Pontina km 30.600, 00040 Pomezia, Rome, Italy The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors
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