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Originally published In Press as doi:10.1074/jbc.M910303199 on March 23, 2000

J. Biol. Chem., Vol. 275, Issue 22, 17086-17093, June 2, 2000
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Purification of the Novel Endonuclease, Hpy188I, and Cloning of Its Restriction-Modification Genes Reveal Evidence of Its Horizontal Transfer to the Helicobacter pylori Genome*

Qing XuDagger , Shawn Stickel§, Richard J. Roberts§, Martin J. BlaserDagger , and Richard D. Morgan§||

From the Dagger  Department of Microbiology and Immunology,  Division of Infectious Diseases, Department of Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee 37232 and § New England Biolabs, Inc., Beverly, Massachusetts 01915

We have isolated a novel restriction endonuclease, Hpy188I, from Helicobacter pylori strain J188. Hpy188I recognizes the unique sequence, TCNGA, and cleaves the DNA between nucleotides N and G in its recognition sequence to generate a one-base 3' overhang. Cloning and sequence analysis of the Hpy188I modification gene in strain J188 reveal that hpy188IM has a 1299-base pair (bp) open reading frame (ORF) encoding a 432-amino acid product. The predicted protein sequence of M.Hpy188I contains conserved motifs typical of aminomethyltransferases, and Western blotting indicates that it is an N-6 adenine methyltransferase. Downstream of hpy188IM is a 513-bp ORF encoding a 170-amino acid product, that has a 41-bp overlap with hpy188IM. The predicted protein sequence from this ORF matches the amino acid sequence obtained from purified Hpy188I, indicating that it encodes the endonuclease. The Hpy188I R-M genes are not present in either strain of H. pylori that has been completely sequenced but are found in two of 11 H. pylori strains tested. The significantly lower G + C content of the Hpy188I R-M genes implies that they have been introduced relatively recently during the evolution of the H. pylori genome.

* This work was supported in part by a Dissertation Enhancement Grant from the Vanderbilt Graduate School, National Institutes of Health Grants GM56534 and DK53707, and a Vanderbilt Cancer Center Core Grant.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequences reported in this paper have been submitted to the GenBankTM/EBI Data Bank with accession numbers AF202061 (the sequence of the hpy188IM-Hpy188IR locus in strain J188), AF215914 (the sequence of the corresponding region in strain A101), AF215915 (that in strain J262), AF215916 (that in strain J178), and AF215917 (that in strain 60190).

|| To whom correspondence should be addressed: New England Biolabs, Inc., 32 Tozer Rd., Beverly, MA 01915. Tel.: 978-927-5054; Fax: 978-921-1350; E-mail: morgan@neb.com.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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