Effect of growth phase and acid shock on Helicobacter pylori cagA expression.
Helicobacter pylori strains possessing cagA are associated with peptic ulceration. To understand the regulation of expression of cagA, picB, associated with interleukin-8 induction, and ureA, encoding the small urease subunit, we created gene fusions of cagA, ureA, and picB of strain 3401, using a promoterless reporter (xylE). Expression of XylE after growth in broth culture revealed that basal levels of expression of cagA and urea in H. pylori were substantially greater than for picB. For cagA expression in stationary-phase cells, brief exposure to acid pH caused a significant increase in xylE expression compared with neutral pH. In contrast, expression of xylE in urea or picB decreased after parallel exposure to acid pH (pH 7 > 6 > 5 > 4), regardless of the growth phase. Expression of the CagA protein varied with growth phase and pH exposure in parallel with the observed transcriptional variation. The concentration of CagA in a cell membrane-enriched fraction after growth at pH 6 was significantly higher than after growth at pH 5 or 7. We conclude that the promoterless reporter xylE is useful for studying the regulation of gene expression in H. pylori and that regulation of CagA production occurs mainly at the transcriptional level.
Karita
M
M
Department of Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-2605, USA.
Tummuru
M K
MK
Wirth
H P
HP
Blaser
M J
MJ
D000942:Antigens, Bacterial; D001426:Bacterial Proteins / Q000235:genetics*; D003001:Cloning, Molecular; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D016480:Helicobacter pylori / Q000235:genetics* / Q000254:growth & development / Q000502:physiology; D006863:Hydrogen-Ion Concentration; D015854:Up-Regulation; D014510:Urease / Q000235:genetics
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C080866:cagA protein, Helicobacter pylori; D014510:Urease