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PMID: 2277628 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

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

Molecular & general genetics : MGG ·Vol. 224 ·No. 1 ·1990-10-00 ·Pages 101-10

Labbé D, Höltke HJ, Lau PC

Abstract

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 EcoRI-PstI fragment localizes the M. NlaIII gene to a 334 codon open reading frame (ORF) and identifies, 468 bp 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 wellknown 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.

Related Genes
M.NlaII
MeSH Terms
Amino Acid Sequence Base Sequence Cloning, Molecular DNA-Cytosine Methylases/genetics,metabolism Genes, Bacterial Molecular Sequence Data Neisseria/enzymology,genetics Open Reading Frames Restriction Mapping Sequence Homology, Nucleic Acid Site-Specific DNA-Methyltransferase (Adenine-Specific)/genetics,metabolism
Chemicals
DNA-Cytosine Methylases Site-Specific DNA-Methyltransferase (Adenine-Specific)
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Labbé D
Biotechnology Research Institute, National Research Council of Canada, Montreal, Quebec.
Höltke H J
Lau P C
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00259456
Published
1990-10-00
Pages
101-10
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00259456
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