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Journal of Bacteriology, March 2007, p. 2426-2434, Vol. 189, No. 6
0021-9193/07/$08.00+0     doi:10.1128/JB.01492-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

The HP0165-HP0166 Two-Component System (ArsRS) Regulates Acid-Induced Expression of HP1186 {alpha}-Carbonic Anhydrase in Helicobacter pylori by Activating the pH-Dependent Promoter{triangledown}

Yi Wen,* Jing Feng, David R. Scott, Elizabeth A. Marcus, and George Sachs

The Membrane Biology Laboratory, Department of Physiology, David Geffen School of Medicine at UCLA, and VA Greater Los Angeles Healthcare System, Los Angeles, California 90073

Received 21 September 2006/ Accepted 3 January 2007

The periplasmic {alpha}-carbonic anhydrase of Helicobacter pylori is essential for buffering the periplasm at acidic pH. This enzyme is an integral component of the acid acclimation response that allows this neutralophile to colonize the stomach. Transcription of the HP1186 {alpha}-carbonic anhydrase gene is upregulated in response to low environmental pH. A binding site for the HP0166 response regulator (ArsR) has been identified in the promoter region of the HP1186 gene. To investigate the mechanism that regulates the expression of HP1186 in response to low pH and the role of the HP0165-HP0166 two-component system (ArsRS) in this acid-inducible regulation, Northern blot analysis was performed with RNAs isolated from two different wild-type H. pylori strains (26695 and 43504) and mutants with HP0165 histidine kinase (ArsS) deletions, after exposure to either neutral pH or low pH (pH 4.5). ArsS-dependent upregulation of HP1186 {alpha}-carbonic anhydrase in response to low pH was found in both strains. Western blot analysis of H. pylori membrane proteins confirmed the regulatory role of ArsS in HP1186 expression in response to low pH. Analysis of the HP1186 promoter region revealed two possible transcription start points (TSP1 and TSP2) located 43 and 11 bp 5' of the ATG start codon, respectively, suggesting that there are two promoters transcribing the HP1186 gene. Quantitative primer extension analysis showed that the promoter from TSP1 (43 bp 5' of the ATG start codon) is a pH-dependent promoter and is regulated by ArsRS in combating environmental acidity, whereas the promoter from TSP2 may be responsible for control of the basal transcription of HP1186 {alpha}-carbonic anhydrase.

* Corresponding author. Mailing address: 11301 Wilshire Blvd., Bldg. 113, Rm. 324, Los Angeles, CA 90073. Phone: (310) 268-4672. Fax: (310) 312-9478. E-mail: ywen{at}ucla.edu.

{triangledown} Published ahead of print on 12 January 2007.

Journal of Bacteriology, March 2007, p. 2426-2434, Vol. 189, No. 6
0021-9193/07/$08.00+0     doi:10.1128/JB.01492-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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