Tl. Norris et al., EXPRESSION AND TRANSCRIPTIONAL CONTROL OF THE SALMONELLA-TYPHIMURIUM LPF FIMBRIAL OPERON BY PHASE VARIATION, Molecular microbiology, 29(1), 1998, pp. 311-320
The lpf operon mediates adhesion of Salmonella typhimurium to murine P
eyer's patches. To investigate expression of this virulence factor, a
transcriptional fusion between the lpf operon and the genes lacZYA of
Escherichia coil was constructed and introduced into the S. typhimuriu
m chromosome. The resulting strain yielded both Lac(+) and Lac(-) colo
ny phenotypes. Alternation between Lac(+) (phase ON) and Lac(-) (phase
OFF) phenotypes occurred by a heritable phase Variation mechanism, as
inoculation of broth cultures with bacteria picked from a Laci colony
gave rise to a considerably higher proportion of Lac(+) colonies than
inoculation with bacteria picked from a Lac(-) colony. During growth
in vitro, phase transition from ON to OFF and from OFF to ON occurred
at rates of 6.8x 10(-3) and 2.4 x10(-4) events per cell per generation
respectively. In a murine intestinal organ culture model, selection f
or the ON expression state occurred when attached bacteria were recove
red from Peyer's patches, suggesting that Lac phase variation correlat
ed with expression of lpf mediated adherence. Selection for either the
ON or the OFF expression state of the lpf operon in vivo was studied
in mice immunized with either GST or GST-LpfA fusion protein. A strong
selection against phase ON cells occurred only in animals immunized w
ith GST-LpfA. The ability of animals immunized with GST-LpfA to distin
guish between phase ON and phase OFF bacteria provided evidence for th
e presence of LpfA fimbrial protein in phase ON cells and for its expo
sure to the immune system.