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Published online on June 5, 2006, 10.1073/pnas.0601754103
PNAS | June 13, 2006 | vol. 103 | no. 24 | 9280-9285


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BIOLOGICAL SCIENCES / MICROBIOLOGY
A Vibrio cholerae protease needed for killing of Caenorhabditis elegans has a role in protection from natural predator grazing

Karolis Vaitkevicius*, Barbro Lindmark*, Gangwei Ou{dagger}, Tianyan Song*, Claudia Toma{ddagger}, Masaaki Iwanaga{ddagger}, Jun Zhu§, Agneta Andersson, Marie-Louise Hammarström{dagger}, Simon Tuck||, and Sun Nyunt Wai*,**

Departments of *Molecular Biology and Ecology and Environmental Science, and ||Umeå Center for Molecular Pathogenesis, Umeå University, SE-90187 Umeå, Sweden; {dagger}Department of Clinical Microbiology, Division of Immunology, Umeå University, SE-90185 Umeå, Sweden; {ddagger}Division of Bacterial Pathogenesis, Graduate School of Medicine, University of the Ryukyus, Okinawa 903-0215, Japan; and §Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104

Edited by Emil C. Gotschlich, The Rockefeller University, New York, NY, and approved May 2, 2006 (received for review March 2, 2006)

Vibrio cholerae is the causal bacterium of the diarrheal disease cholera, and its growth and survival are thought to be curtailed by bacteriovorous predators, e.g., ciliates and flagellates. We explored Caenorhabditis elegans as a test organism after finding that V. cholerae can cause lethal infection of this nematode. By reverse genetics we identified an extracellular protease, the previously uncharacterized PrtV protein, as being necessary for killing. The killing effect is associated with the colonization of bacteria within the Caenorhabditis elegans intestine. We also show that PrtV is essential for V. cholerae in the bacterial survival from grazing by the flagellate Cafeteria roenbergensis and the ciliate Tetrahymena pyriformis. The PrtV protein appears to have an indirect role in the interaction of V. cholerae with mammalian host cells as judged from tests with tight monolayers of human intestinal epithelial cells. Our results demonstrate a key role for PrtV in V. cholerae interaction with grazing predators, and we establish Caenorhabditis elegans as a convenient organism for identification of V. cholerae factors involved in host interactions and environmental persistence.

cholera | host interactions | environmental persistence

Author contributions: K.V., A.A., M.-L.H., S.T., and S.N.W. designed research; K.V., B.L., G.O., J.Z., S.T., and S.N.W. performed research; K.V., J.Z., A.A., M.-L.H., S.T., and S.N.W. analyzed data; T.S., C.T., M.I., J.Z., A.A., M.-L.H., and S.T. contributed new reagents/analytic tools; and S.N.W. wrote the paper.

Conflict of interest statement: No conflicts declared.

This paper was submitted directly (Track II) to the PNAS office.

**To whom correspondence should be addressed. E-mail: sun.nyunt.wai{at}molbiol.umu.se

© 2006 by The National Academy of Sciences of the USA


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