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Nucleic Acids Research, 1992, Vol. 20, No. 19 5047-5052
© 1992

MOLECULAR BIOLOGY

Identification of the CTAG-recognizing restriction-modification systems MthZI and MthFI from Methanobacterium thermoformicicum and characterization of the plasmid-encoded mthZIM gene

Jörk Nölling and Willem M.de Vos*

Bacterial Genetics Group, Department of Microbiology, Wageningen Agricultural University Hesselink van Suchtelenweg 4, 6703 CT Wageningen, The Netherlands

* To whom correspondence should be addressed

Received July 14, 1992. Revised September 9, 1992. Accepted September 9, 1992.

Two CTAG-recognlzing restriction and modification (R/M) systems, designated MthZI and MthFI, were identified in the thermophllic archaeon Methanobacterium thermoformicicum strains Z-245 and FTF, respectively. Further analysis revealed that the methyltransferase (MTase) genes are plasmid-located in both strains. The plasmid pFZ1-encoded mthZIM gene of strain Z-245 was further characterized by subclonlng and expression studies in Escherlchla coli followed by nucleotide sequence analysis. The mthZIM gene Is 1065 bp in size and may code for a protein of 355 amlno acids (Mr 42,476 Da). The deduced amino acid sequence of the M.MthZI enzyme shares substantial similarity with four distinct regions from several m4C-and m6A-MTases, and contains the TSPPY motif that is so far only found in m4C-MTases. Partially overlapping with the mthZIM gene and in reverse orientation, an additional ORF was identified with a size of 606 bp potentially coding for a protein of 202 amino acids (Mr 23.710 Da). This ORF is suggested to encode the corresponding endonuclease R.MthZI.


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