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Cloning and characterization of two tandemly arranged DNA methyltransferase genes of Neisseria lactamica: An adenine-specific M.NlaIII and a cytosine-type methylase

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Summary

The gene encoding the Neisseria lactamica III DNA methyltransferase (M.NlaIII) which recognizes the sequence CATG has been cloned and expressed in Escherichia coli. DNA sequencing of a 3.125 kb EcoR1-Pst1 fragment localizes the M.NlaIII gene to a 334 codon open reading frame (ORF) and identifies, 468 by downstream, a second ORF of 313 amino acids, which is referred to as M.NlaX. Both proteins are detectable in the E. coli coupled in vitro transcription-translation system; they are apparently expressed from separate N. lactamica promoters. The N-terminal half of the previously characterized M.FokI, which methylates adenine in one of the DNA strands with its asymmetric recognition sequence (GGATG), is found to have 41% sequence identity and a further 11.7% sequence similarity with M.NlaIII. Among the conserved amino acids is the well-known DPPY sequence motif. With one exception, analysis of the nucleotides coding for the DP dipeptide in all known DPPY sequences shows the presence of an inherent DNA adenine methylation (dam) recognition site of GATC. A low level of expression of M.NlaX in E. coli prevents the elucidation of its sequence recognition specificity. Sequence analysis of M.NlaX shows that it is closely related to the group of monospecific 5-methylcytosine DNA methyltransferases (M.EcoRII, Dcm, M.HpaII and M.HhaI) which all have a modified cytosine at the second position of the recognition sequences. Both M.EcoRII and Dcm amino acid sequences are about 50% identical with M.NlaX; a considerable degree of sequence identity is found in the so-called variable region which is believed to be responsible for sequence recognition specificity. M.NlaX is probably the counterpart to the E. coli Dcm in N. lactamica.

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Authors and Affiliations

  1. Biotechnology Research Institute, National Research Council of Canada, 6100 Royalmount Ave, H4P 2R2, Montreal, Quebec, Canada

    Diane Labbé & Peter C. K. Lau

  2. Department of Molecular Biology, Boehringer Mannheim GmbH, Biochemical Research Centre, Nonnenwald 2, D-8122, Penzberg, Federal Republic of Germany

    H. Joachim Höltke

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  1. Diane Labbé
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  2. H. Joachim Höltke
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  3. Peter C. K. Lau
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Communicated by W. Goebel

Issued as NRCC publication number 31454

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Labbé, D., Höltke, H.J. & Lau, P.C.K. Cloning and characterization of two tandemly arranged DNA methyltransferase genes of Neisseria lactamica: An adenine-specific M.NlaIII and a cytosine-type methylase. Molec. Gen. Genet. 224, 101–110 (1990). https://doi.org/10.1007/BF00259456

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