Open AccessCloning, sequence analysis and heterologous expression of the DNA adenine‐( N 6 ) methyltransferase from the human pathogen Actinobacillus actinomycetemcomitans
Author(s) -
Eberhard Jörg,
Oza Javin,
Reich Norbert O
Publication year - 2001
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2001.tb10525.x
Subject(s) - cloning (programming) , actinobacillus , heterologous expression , heterologous , biology , pathogen , dna , molecular cloning , sequence analysis , methyltransferase , microbiology and biotechnology , human pathogen , genetics , chemistry , recombinant dna , peptide sequence , gene , bacteria , methylation , computer science , programming language
We cloned and sequenced the DNA adenine‐ N 6 methyltransferase gene of the human pathogen Actinobacillus actinomycetemcomitans (M. Aac DAM). Restriction digestion shows that the enzyme methylates adenine in the sequence GATC. Expression of the enzyme in a DAM − background shows in vivo activity. A PSI‐BLAST search revealed that M. Aac DAM is most related to M. Hin dIV, M. Eco DAM, M. Sty DAM, and M. Sma II. The ClustalW alignment shows highly conserved regions in the enzyme characteristic for type a MTases. Phylogenetic tree analysis shows a cluster of enzymes recognizing the sequence GATC, within a branch of orphan MTases harboring M. Aac DAM. The cloning and sequencing of this first methyltransferase gene described for A. actinomycetemcomitans open the path for studies on the potential regulatory impact of DNA methylation on gene regulation and virulence in this organism.