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Microbiology 149 (2003), 1883-1891; DOI  10.1099/mic.0.26086-0
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Microbiology 149 (2003), 1883-1891; DOI  10.1099/mic.0.26086-0
© 2003 Society for General Microbiology

Haemagglutinin/protease expression and mucin gel penetration in El Tor biotype Vibrio cholerae

Anisia J. Silva2, Kim Pham2 and Jorge A. Benitez1

1 Morehouse School of Medicine Department of Microbiology, Biochemistry and Immunology, 720 Westview Dr, SW, Atlanta, GA 30310-1495, USA
2 California State University College of Science and Mathematics, Department Biology, 2555 East San Ramon Avenue, Fresno, CA 93740, USA

Correspondence
Jorge A. Benitez
jbenitez{at}msm.edu

Vibrio cholerae of both biotypes produce a soluble Zn2+-dependent metalloprotease: haemagglutinin/protease (Hap), encoded by hapA. Hap has been shown to have mucinolytic and cytotoxic activity. These activities are likely to play an important role in the pathogenesis of cholera and the reactogenicity of attenuated vaccine strains. Production of Hap requires transcriptional activation by the HapR regulator and is repressed by glucose. The present study shows that mucin purified from two sources, bile salts, and growth at 37 °C enhanced Hap protease production. Analysis of hapA and hapR promoter fusions with the lacZ gene showed both promoters to be activated in a cell-density-dependent pattern. Glucose repressed and mucin induced the hapA promoter by a HapR-independent mechanism. Bile had no effect on either hapR or hapA promoter activity. Expression of hapA was required for vibrios to translocate through a mucin-containing gel. These results suggest Hap to play an important role in cholera pathogenesis by promoting mucin gel penetration, detachment and spreading of infection along the gastrointestinal tract.

Abbreviations: CRP, cAMP receptor protein; PS, porcine stomach; SMG, submaxillary gland; TCP, toxin co-regulated pilus




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