1887

Abstract

The nonadhesive mutant CD11 of El Tor, defective in expression of mannose-sensitive haemagglutinin, lacks a protein when compared with its parent strain. Determination of the amino acid sequence revealed the identity of the protein as the product of VC1929, which is annotated to encode a protein, DctP, involved in the transport of C-dicarboxylates. We cloned the gene in pUC19 vector and expressed it in mutant CD11. Expression of DctP in the resulting complemented strain restored virulence, adhesive and colonizing capabilities, mannose-sensitive haemagglutination (MSHA) and ability to grow in medium containing sialic acid as a sole carbon source. The mutation in CD11 was caused by insertion of an adenine nucleotide in the reading frame of . Recombinant purified DctP protein showed MSHA of human red blood cells, and protected rabbits against infection by The protein was localized in membrane and cell wall fractions. The mutant, recombinant CD11 expressing DctP and parent strains were grown in M9 minimal medium in the presence of various carbohydrates (glucose, malate, fumarate, succinate or -acetylneuraminic acid). The mutant was unable to grow in minimal medium containing -acetylneuraminic acid (sialic acid) as the sole carbon source whereas the recombinant and parent strains utilized all the sugars tested. It is concluded that DctP is a mannose-sensitive haemagglutinin and a virulence factor and is involved in the utilization of sialic acid.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.050245-0
2011-11-01
2019-12-11
Loading full text...

Full text loading...

/deliver/fulltext/micro/157/11/3180.html?itemId=/content/journal/micro/10.1099/mic.0.050245-0&mimeType=html&fmt=ahah

References

  1. Almagro-Moreno S. , Boyd E. F. . ( 2009; ). Sialic acid catabolism confers a competitive advantage to pathogenic Vibrio cholerae in the mouse intestine. . Infect Immun 77:, 3807–3816. [CrossRef] [PubMed]
    [Google Scholar]
  2. Attridge S. R. , Manning P. A. , Holmgren J. , Jonson G. . ( 1996; ). Relative significance of mannose-sensitive hemagglutinin and toxin-coregulated pili in colonization of infant mice by Vibrio cholerae El Tor. . Infect Immun 64:, 3369–3373.[PubMed]
    [Google Scholar]
  3. Bhattacharjee J. W. , Srivastava B. S. . ( 1978; ). Mannose-sensitive haemagglutinins in adherence of Vibrio cholerae eltor to intestine. . J Gen Microbiol 107:, 407–410.[PubMed] [CrossRef]
    [Google Scholar]
  4. Bradford M. M. . ( 1976; ). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. . Anal Biochem 72:, 248–254. [CrossRef] [PubMed]
    [Google Scholar]
  5. Burrows M. R. , Sellwood R. , Gibbons R. A. . ( 1976; ). Haemagglutinating and adhesive properties associated with the K99 antigen of bovine strains of Escherichia coli . . J Gen Microbiol 96:, 269–275.[PubMed] [CrossRef]
    [Google Scholar]
  6. Elwell L. P. , Shipley P. L. . ( 1980; ). Plasmid-mediated factors associated with virulence of bacteria to animals. . Annu Rev Microbiol 34:, 465–496. [CrossRef] [PubMed]
    [Google Scholar]
  7. Field M. , Rao M. C. , Chang E. B. . ( 1989; ). Intestinal electrolyte transport and diarrheal disease (2). . N Engl J Med 321:, 879–883. [CrossRef] [PubMed]
    [Google Scholar]
  8. Finkelstein R. A. . ( 1962; ). Vibriocidal antibody inhibition (VAI) analysis: a technique for the identification of the predominant vibriocidal antibodies in serum and for the detection and identification of Vibrio cholerae antigens. . J Immunol 89:, 264–271.
    [Google Scholar]
  9. Galen J. E. , Ketley J. M. , Fasano A. , Richardson S. H. , Wasserman S. S. , Kaper J. B. . ( 1992; ). Role of Vibrio cholerae neuraminidase in the function of cholera toxin. . Infect Immun 60:, 406–415.[PubMed]
    [Google Scholar]
  10. Gupta R. K. , Srivastava B. S. , Srivastava R. . ( 2010; ). Comparative expression analysis of rpf-like genes of Mycobacterium tuberculosis H37Rv under different physiological stress and growth conditions. . Microbiology 156:, 2714–2722. [CrossRef] [PubMed]
    [Google Scholar]
  11. Hanahan D. . ( 1983; ). Studies on transformation of Escherichia coli with plasmids. . J Mol Biol 166:, 557–580. [CrossRef] [PubMed]
    [Google Scholar]
  12. Heidelberg J. F. , Eisen J. A. , Nelson W. C. , Clayton R. A. , Gwinn M. L. , Dodson R. J. , Haft D. H. , Hickey E. K. , Peterson J. D. et al. ( 2000; ). DNA sequence of both chromosomes of the cholera pathogen Vibrio cholerae . . Nature 406:, 477–483. [CrossRef] [PubMed]
    [Google Scholar]
  13. Jacob A. , Sinha V. B. , Sahib M. K. , Srivastava R. , Kaper J. B. , Srivastava B. S. . ( 1993; ). Identification of a 33 kDa antigen associated with an adhesive and colonizing strain of Vibrio cholerae El Tor and its role in protection. . Vaccine 11:, 376–382. [CrossRef] [PubMed]
    [Google Scholar]
  14. Janausch I. G. , Zientz E. , Tran Q. H. , Kröger A. , Unden G. . ( 2002; ). C4-dicarboxylate carriers and sensors in bacteria. . Biochim Biophys Acta 1553:, 39–56. [CrossRef] [PubMed]
    [Google Scholar]
  15. Jones G. W. , Freter R. . ( 1976; ). Adhesive properties of Vibrio cholerae: nature of the interaction with isolated rabbit brush border membranes and human erythrocytes. . Infect Immun 14:, 240–245.[PubMed]
    [Google Scholar]
  16. Jones G. W. , Rutter J. M. . ( 1974; ). The association of K88 antigen with haemagglutinating activity in porcine strains of Escherichia coli . . J Gen Microbiol 84:, 135–144.[PubMed] [CrossRef]
    [Google Scholar]
  17. Jonson G. , Holmgren J. , Svennerholm A. M. . ( 1991; ). Identification of a mannose-binding pilus on Vibrio cholerae El Tor. . Microb Pathog 11:, 433–441. [CrossRef] [PubMed]
    [Google Scholar]
  18. Jonson G. , Lebens M. , Holmgren J. . ( 1994; ). Cloning and sequencing of Vibrio cholerae mannose-sensitive haemagglutinin pilin gene: localization of mshA within a cluster of type 4 pilin genes. . Mol Microbiol 13:, 109–118. [CrossRef] [PubMed]
    [Google Scholar]
  19. Kaper J. B. , Morris J. G. Jr , Levine M. M. . ( 1995; ). Cholera. . Clin Microbiol Rev 8:, 48–86.[PubMed]
    [Google Scholar]
  20. Koransky J. R. , Scales R. W. , Kraus S. J. . ( 1975; ). Bacterial hemagglutination by Neisseria gonorrhoeae . . Infect Immun 12:, 495–498.[PubMed]
    [Google Scholar]
  21. Larkin M. A. , Blackshields G. , Brown N. P. , Chenna R. , McGettigan P. A. , McWilliam H. , Valentin F. , Wallace I. M. , Wilm A. et al. ( 2007; ). clustal w and clustal_x version 2.0. . Bioinformatics 23:, 2947–2948. [CrossRef] [PubMed]
    [Google Scholar]
  22. Lee B. Y. , Hefta S. A. , Brennan P. J. . ( 1992; ). Characterization of the major membrane protein of virulent Mycobacterium tuberculosis. . Infect Immun 60:, 2066–2074.[PubMed]
    [Google Scholar]
  23. Marcus H. , Ketley J. M. , Kaper J. B. , Holmes R. K. . ( 1990; ). Effects of DNase production, plasmid size, and restriction barriers on transformation of Vibrio cholerae by electroporation and osmotic shock. . FEMS Microbiol Lett 68:, 149–154. [CrossRef] [PubMed]
    [Google Scholar]
  24. Miller J. H. . ( 1972; ). Experiments in Molecular Genetics. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory Press;.
    [Google Scholar]
  25. Ofek I. , Mirelman D. , Sharon N. . ( 1977; ). Adherence of Escherichia coli to human mucosal cells mediated by mannose receptors. . Nature 265:, 623–625. [CrossRef] [PubMed]
    [Google Scholar]
  26. Sambrook J. , Fritsch E. , Maniatis T. . ( 1989; ). Molecular Cloning: a Laboratory Manual. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory Press;.
    [Google Scholar]
  27. Severi E. , Hood D. W. , Thomas G. H. . ( 2007; ). Sialic acid utilization by bacterial pathogens. . Microbiology 153:, 2817–2822. [CrossRef] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.050245-0
Loading
/content/journal/micro/10.1099/mic.0.050245-0
Loading

Data & Media loading...

Supplements

Supplementary figure 

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error