Phase variation in Bordetella pertussis is accompanied by changes in DNA modification.
Pathogenic strains of Bordetella pertussis tend to undergo a phase variation process when propagated in vitro. The phase variants do not express part or all of the virulence factors of the pathogenic strain and are phenotypically stable. We have previously shown that variation involves a non-reversible, non-random process. In an attempt to characterize the molecular changes accompanying phase variation, chromosomal DNA, isolated from B. pertussis and its variants, was digested with a variety of restriction enzymes followed by agarose gel electrophoresis. While variant DNA was digested by all tested enzymes, pathogenic strain DNA was not digested by part of the enzymes, thus suggesting modification of the DNA at specific sites. DNA isolated from reversible, growth medium induced variants, demonstrated sensitivity to digestion identical to that of spontaneous, stable variants. Analysis of the restriction sequences of all the enzymes, which did not digest DNA from pathogenic strains, failed to reveal any common sequence known to be affected by methylation. HPLC and nearest-neighbor analysis showed a 2-fold increase in the level of DNA methylation in the pathogenic strain. It was concluded that (a) the chromosomal DNA in virulent strains of B. pertussis is protected against enzymatic digestion by an as yet unknown modification and (b) variation in B. pertussis may be caused by changes in the modification of the DNA rather than by mutation.
Goldman
S
S
Department of Microbiology, Faculty of Life Sciences, Tel Aviv University, Israel.
Navon
Y
Y
Fish
F
F
D001483:Base Sequence; D001886:Bordetella pertussis / Q000235:genetics* / Q000472:pathogenicity; D002851:Chromatography, High Pressure Liquid; D002876:Chromosomes, Bacterial / Q000032:analysis; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D014644:Genetic Variation; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D014774:Virulence
D004269:DNA, Bacterial