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Infection and Immunity, February 2000, p. 470-477, Vol. 68, No. 2
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Expression of the Helicobacter pylori
ureI Gene Is Required for Acidic pH Activation of
Cytoplasmic Urease
David R.
Scott,1,*
Elizabeth A.
Marcus,1
David L.
Weeks,1
Adrian
Lee,2
Klaus
Melchers,3 and
George
Sachs1
Department of Physiology, University of
California at Los Angeles, and VAGLAHS, Los Angeles,
California1; University of New
South Wales, Sydney, Australia2; and
Byk Gulden, Konstanz, Germany3
Received 9 August 1999/Returned for modification 29 September
1999/Accepted 3 November 1999
ureI encodes an integral cytoplasmic membrane protein.
It is present in the urease gene cluster of Helicobacter
pylori and is essential for infection and acid survival, but its
role is unknown. To determine the function of UreI protein, we produced H. pylori ureI deletion mutants and measured the pH
dependence of urease activity of intact and lysed bacteria and the
effect of urea on the membrane potential. We also determined
ureI expression, urease activity, and the effect of urea on
membrane potential of several gastric and nongastric
Helicobacter species. ureI was found to be
present in the genome of the gastric Helicobacter species
and absent in the nongastric Helicobacter species studied, as determined by PCR. Likewise, Western blot analysis confirmed that
UreI was expressed only in the gastric Helicobacter
species. When UreI is present, acidic medium pH activation of
cytoplasmic urease is found, and urea addition increases membrane
potential at acidic pH. The addition of a low concentration of
detergent raised urease activity of intact bacteria at neutral pH to
that of their homogenates, showing that urease activity was membrane limited. No acidic pH activation or urea induced membrane potential changes were found in the nongastric Helicobacter species.
The ureI gene product is probably a pH activated urea
transporter or perhaps regulates such a transporter as a function of
periplasmic pH.
*
Corresponding author. Mailing address: VAGLAHS, Bldg.
113, Rm. 324, 11301 Wilshire Blvd., Los Angeles, CA 90073. Phone: (310) 268-4672. Fax: (310) 312-9478. E-mail: dscott{at}ucla.edu.
Infection and Immunity, February 2000, p. 470-477, Vol. 68, No. 2
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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