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A bacteriophage encoding a pathogenicity island, a type-IV pilus and a phage receptor in cholera bacteria

Nature volume 399, pages 375–379 (1999)Cite this article

Abstract

The virulence properties of many pathogenic bacteria are due to proteins encoded by large gene clusters called pathogenicity islands1,2, which are found in a variety of human pathogens including Escherichia coli, Salmonella, Shigella, Yersinia, Helicobacter pylori, Vibrio cholerae, and animal and plant pathogens such as Dichelobacter nodosus and Pseudomonas syringae1,2,3. Although the presence of pathogenicity islands is a prerequisite for many bacterial diseases, little is known about their origins or mechanism of transfer into the bacterium. The bacterial agent of epidemic cholera, Vibrio cholerae , contains a bacteriophage known as cholera-toxin phage (CTXφ)4, which encodes the cholera toxin, and a large pathogenicity island called the VPI (for V. cholerae pathogenicity island)5 which itself encodes a toxin-coregulated pilus that functions as a colonization factor6 and as a CTXφ receptor4. We have now identified the VPI pathogenicity island as the genome of another filamentous bacteriophage, VPIφ. We show that VPIφ is transferred between V. cholerae strains and provide evidence that the TcpA subunit of the toxin-coregulated type IV pilus is in fact a coat protein of VPIφ. Our results are the first description of a phage that encodes a receptor for another phage and of a virus–virus interaction that is necessary for bacterial pathogenicity.

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Figure 1: Diagram of the VPI.
Figure 2: PCR products obtained from either whole-cell overnight cultures or cell-free phage (φ) preparations of wild-type strain N16961 and TCP2.
Figure 3: Southern blot of phage DNA preparations from DK238 and CVD110.
Figure 4: Analysis of the VPIφ replicative form.
Figure 5: Electron micrographs of VPIφ.
Figure 6: PCR products of immunoprecipitated phage preparations from strains 395 and TCP2, following incubation with and without anti-TCP antibodies (Ab), demonstrate specificity of anti-TCP antibody for VPIφ.
Figure 7: PCR analysis of VPI genes from a tcpA mutant and mutant complemented with tcpA.

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Acknowledgements

We thank our colleagues at the University of Maryland for their support, particularly D. Hone for discussions, T. Agin, J. Michalski and M. Boyanapalli for technical assistance, and R. T. Milanich for help with prepAring the figures; and R. Taylor for RT4032, pRT198 and anti-peptide 6 TcpA antibody. This work was supported by grants from the NIH and Department of Veterans Affairs. D.K.R.K. is a recipient of a Burroughs Wellcome Fund Career Award in the Biomedical Sciences.

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Authors and Affiliations

  1. Center for Vaccine Development, Baltimore, 21201, Maryland, USA

    David K. R. Karaolis, Sita Somara, David R. Maneval Jr & James B. Kaper

  2. Division of Hospital Epidemiology, Baltimore, 21201, Maryland, USA

    David K. R. Karaolis

  3. Departments of Pathology, Baltimore, 21201, Maryland, USA

    Judith A. Johnson

  4. Departments of Microbiology and Immunology University of Maryland School of Medicine, Baltimore, 21201, Maryland, USA

    James B. Kaper

  5. Department of Veterans Affairs, Maryland Health Care System, Baltimore, 21201 , Maryland, USA

    Judith A. Johnson

Authors
  1. David K. R. Karaolis
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  2. Sita Somara
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  3. David R. Maneval Jr
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  4. Judith A. Johnson
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  5. James B. Kaper
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Correspondence to David K. R. Karaolis.

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Karaolis, D., Somara, S., Maneval, D. et al. A bacteriophage encoding a pathogenicity island, a type-IV pilus and a phage receptor in cholera bacteria. Nature 399, 375–379 (1999). https://doi.org/10.1038/20715

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