Functional analysis of putative restriction-modification system genes in the Helicobacter pylori J99 genome.
Helicobacter pylori is a gram-negative bacterium, which colonizes the gastric mucosa of humans and is implicated in a wide range of gastroduodenal diseases. The genomic sequences of two H.pylori strains, 26695 and J99, have been published recently. About two dozen potential restriction-modification (R-M) systems have been annotated in both genomes, which is far above the average number of R-M systems in other sequenced genomes. Here we describe a functional analysis of the 16 putative Type II R-M systems in the H. pylori J99 genome. To express potentially toxic endonuclease genes, a unique vector was constructed, which features repression and antisense transcription as dual control elements. To determine the methylation activities of putative DNA methyltransferases, we developed polyclonal antibodies able to detect DNA containing N6-methyladenine or N4-methylcytosine. We found that <30% of the potential Type II R-M systems in H.pylori J99 strain were fully functional, displaying both endonuclease and methyltransferase activities. Helicobacter pylori may maintain a variety of functional R-M systems, which are believed to be a primitive bacterial 'immune' system, by alternatively turning on/off a subset of numerous R-M systems.
Kong
H
H
New England Biolabs, 32 Tozer Road, Beverly, MA 01915, USA. kong@neb.com
Lin
L F
LF
Porter
N
N
Stickel
S
S
Byrd
D
D
Posfai
J
J
Roberts
R J
RJ
D000225:Adenine / Q000031:analogs & derivatives* / Q000276:immunology / Q000378:metabolism; D000906:Antibodies / Q000276:immunology; D003001:Cloning, Molecular; D019295:Computational Biology; D003596:Cytosine / Q000031:analogs & derivatives* / Q000276:immunology / Q000378:metabolism; D019175:DNA Methylation; D015254:DNA Modification Methylases / Q000235:genetics* / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000235:genetics* / Q000378:metabolism; D005798:Genes, Bacterial / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D016680:Genome, Bacterial; D016480:Helicobacter pylori / Q000201:enzymology* / Q000235:genetics*; D016366:Open Reading Frames / Q000235:genetics
D000225:Adenine; D000906:Antibodies; D003596:Cytosine; D015254:DNA Modification Methylases; D015252:Deoxyribonucleases, Type II Site-Specific; C039052:N(4)-methylcytosine; C000730567:6-methyladenine