1887

Abstract

To survive and multiply in different environments, has to co-ordinately regulate the expression of genes involved in adaptive responses. In many pathogens, adaptive responses, including pathogenic responses, are regulated by two-component regulator (TCR) systems. It is likely that members of a TCR family play a role in the regulation of processes involved in intestinal colonization, and therefore pathogenesis, in . We have identified and characterized a TCR system of : this system is a homologue of PhoBR. The presence of a putative Pho box suggests that the operon is regulated by inorganic phosphate levels. The and genes are organized the same way as in . Mutation of the gene affected the expression of the putative Pho regulon, including PhoA, but did not affect the production of cholera toxin. mutants are less able to colonize rabbit intestine than wild-type . The addition of inorganic phosphate at a high concentration to the inoculum only partially restored the ability of the mutants to colonize the intestine, suggesting that the Pho regulon may not be regulated by inorganic phosphate levels alone.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-145-9-2463
1999-09-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/145/9/1452463a.html?itemId=/content/journal/micro/10.1099/00221287-145-9-2463&mimeType=html&fmt=ahah

References

  1. Altschul S. F., Gish W., Miller, M., Myers, E. W., Lipman D. J. 1990; Basic local alignment search tool. J Mol Biol 215:403–410 [CrossRef]
    [Google Scholar]
  2. Aoyama T., Takanami M., Makino K., Oka A. 1991; Cross-talk between the virulence and phosphate regulons of Agrobacterium tumefaciens caused by an unusual interaction of the transcriptional activator with a regulatory DNA element. Mol Gen Genet 227:385–390 [CrossRef]
    [Google Scholar]
  3. Ausubel F., Brent R., Kingston R. E., Moore D. D., Seidman J. G., Smith J. A., Struhl K.editors 1990 Current Protocols in Molecular Biology New York: Wiley Interscience;
    [Google Scholar]
  4. Benitez J. A., Spelbrink R. G., Silva A., Phillips, T. E., Stanley C. M., Boesman-Finkelstein M., Finkelstein R. A. 1997; Adherence of Vibrio cholerae to culture differentiated human intestinal cells: an in vitro colonization model. Infect Immun 65:3474–3477
    [Google Scholar]
  5. Bennish M. L. 1994; Cholera: pathophysiology, clinical features, and treatment. In Vibrio Cholerae and Cholera: Molecular to Global Perspectives pp 229–255Edited by Wachsmuth I. K., Blake P. A., Olsvik O. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  6. Bracha M., Yagil E. 1973; A new type of alkaline phosphatase-negative mutants in Escherichia coli.. Mol Gen Genet 122:53–60 [CrossRef]
    [Google Scholar]
  7. 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]
    [Google Scholar]
  8. Comeau D. E., Ikenaka, K, Tsung K., Inouye M. 1985; Primary characterization of the protein products of the Escherichia coli ompR locus – structure and regulation of synthesis of the OmpR and EnvZ proteins. J Bacteriol 164:578–584
    [Google Scholar]
  9. Daigle F., Fairbrother J. M., Harel J. 1995; Identification of a mutation in the pst–phoU operon that reduces pathogenicity of an Escherichia coli strain causing septicemia in pigs. Infect Immun 63:4924–4927
    [Google Scholar]
  10. DiRita V., Peterson K. M., Mekalanos J. J. 1990; Regulation of cholera toxin synthesis. In The Bacteria: Molecular Basis of Bacterial Pathogenesis pp 339–352Edited by Iglewski B. H., Clark V. L. San Diego, CA: Academic Press;
    [Google Scholar]
  11. Echols H., Garen A., Torriani A. 1961; Genetic control of the repression of alkaline phosphatase in Escherichia coli.. J Mol Biol 3:425–438 [CrossRef]
    [Google Scholar]
  12. Fiore A. E., Michalski J. M., Russel R. G., Sears C. L., Kaper J. B. 1997; Cloning, characterization and chromosomal mapping of a phospholipase (lecithinase) produced by Vibrio cholerae.. Infect Immun 65:3112–3117
    [Google Scholar]
  13. Galen J. E., Levine M. M. 1995; Improved suicide vector for chromosomal mutagenesis in Salmonella typhi. In Abstracts of the 95th General Meeting of the American Society for Microbiology p. 158 Washington, DC: American Society for Microbiology;
    [Google Scholar]
  14. Goldberg M. B., DiRita V. J., Calderwood S. B. 1990; Identification of an iron-regulated virulence determinant in Vibrio cholerae, using TnphoA mutagenesis. Infect Immun 58:55–60
    [Google Scholar]
  15. Gutierrez C., Barondess J., Manoil C., Beckwith J. 1987; The use of transposon TnphoA to detect genes for cell-envelope proteins subject to a common regulatory stimulus – analysis of osmotically regulated genes in Escherichia coli.. J Mol Biol 195:289–297 [CrossRef]
    [Google Scholar]
  16. Haldimann A., Daniels L. L., Wanner B. L. 1998; Use of new methods for construction of tightly regulated arabinose and rhamnose promoter fusions in studies of Escherichia coli phosphate regulon. J Bacteriol 180:1277–1286
    [Google Scholar]
  17. Holmgren J. 1973; Comparison of the tissue receptors for Vibrio cholerae and Escherichia coli enterotoxins by means of ganglioside and natural cholera toxoid. Infect Immun 8:851–859
    [Google Scholar]
  18. Humphreys S. J, King S., Ketley J. M. 1995; A PCR-based approach to identify regulatory genes in Vibrio cholerae. In Abstracts of the 95th Ordinary Meeting of the American Society for Microbiology p 214 B280 Washington, DC: American Society for Microbiology;
    [Google Scholar]
  19. Innis M. A., Gelfand D. H. 1990 PCR Protocols: a Guide to Methods and Applications pp 3–12Edited by Innis M. A., Gelfand D. H., Sninsky J. J., White T. J. San Diego, CA: Academic Press;
    [Google Scholar]
  20. Kay A. B., Bopp C. A., Wells J. G. 1994; Isolation and identification of Vibrio cholerae O1 from fecal specimens. In Vibrio cholerae and Cholera: Molecular to Global Perspectives pp 3–25Edited by Wachsmuth I. K., Blake P. A., Olsvik O. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  21. Ketley J. M., Mitchell T. J., Candy D. C. A., Burdon D. W., Stephen J. 1987; The effects of Clostridium difficile crude toxins and toxin-A on ileal and colonic loops in immune and non-immune rabbits. J Med Microbiol 24:41–52 [CrossRef]
    [Google Scholar]
  22. Ketley J. M., Michalski J., Galen, J., Levine, M. M., Kaper J. B. 1993; Construction of genetically marked Vibrio cholerae O1 vaccine strains. FEMS Microbiol Lett 111:15–21 [CrossRef]
    [Google Scholar]
  23. Kornberg A. 1994; Inorganic polyphosphate: a molecular fossil come to life. In Phosphate in Microorganisms: Cellular and Molecular Biology pp 204–208Edited by Torriani-Gorini A., Yagil E., Silver S. Washington DC: American Society for Microbiology;
    [Google Scholar]
  24. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685 [CrossRef]
    [Google Scholar]
  25. Lee S. H., Angelichio M. J., Mekalanos J. J., Camilli A. 1998; Nucleotide sequence and spatiotemporal expression of the Vibrio cholerae vieSAB genes during infection. J Bacteriol 180:2298–2305
    [Google Scholar]
  26. Lee T. Y., Makino K., Shinagawa, H, Amemura M., Nakata A. 1989; Phosphate regulon in members of the family enterobacteriaceae – comparison of the phoBphoR operons of Escherichia coli, Shigella dysenteriae, and Klebsiella pneumoniae.. J Bacteriol 171:6593–6599
    [Google Scholar]
  27. Levine M. M., Tacket C. O. 1994; Recombinant live cholera vaccines. In Vibrio cholerae and Cholera: Molecular to Global Perspectives pp 395–413Edited by Wachsmuth I. K., Blake P. A., Olsvik O. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  28. Levine M. M., Kaper J. B., Herrington D., Ketley J. M., Losonsky G., Tacket C. O, Tall B., Cryz S. 1988; Safety, immunogenicity and efficacy of recombinant live oral cholera vaccines CVD103 and CVD103-HgR. Lancet ii:467–470
    [Google Scholar]
  29. Libby S. J., Goebel W., Muir, S, Songer G., Heffron F. 1990; Cloning and characterization of a cytotoxin gene from Salmonella typhimurium.. Res Microbiol 141:775–783 [CrossRef]
    [Google Scholar]
  30. McCarter L. L., Silverman M. 1987; Phosphate regulation of gene expression in Vibrio parahaemolyticus.. J Bacteriol 169:3441–3449
    [Google Scholar]
  31. Mahan M., Slauch J. M., Mekalanos J. J. 1993; Selection of bacterial virulence genes that are specifically induced in host tissues. Science 259:686–688 [CrossRef]
    [Google Scholar]
  32. Makino K., Shinagawa H., Amemura M., Nakata A. 1986a; Nucleotide sequence of the phoB gene, the positive regulatory gene for the phosphate regulon of Escherichia coli K-12. J Mol Biol 190:37–44 [CrossRef]
    [Google Scholar]
  33. Makino K., Shinagawa H., Amemura M., Nakata A. 1986b; Nucleotide sequence of the phoR gene, a regulatory gene for the phosphate regulon of Escherichia coli.. J Mol Biol 192:549–556 [CrossRef]
    [Google Scholar]
  34. 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]
    [Google Scholar]
  35. Miller J. H. 1972 A Short Course in Bacterial Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  36. Miller V. L., Mekalanos J. J. 1984; Synthesis of cholera toxin is positively regulated at the transcriptional level by ToxR. Proc Natl Acad Sci USA 81:3471–3475 [CrossRef]
    [Google Scholar]
  37. Miller V. L., Mekalanos J. J. 1988; A novel suicide vector and its use in construction of insertion mutations – osmoregulation of outer-membrane proteins and virulence determinants in Vibrio cholerae requires ToxR. J Bacteriol 170:2575–2583
    [Google Scholar]
  38. Miller V. L., Taylor R. K., Mekalanos J. J. 1987; Cholera toxin transcriptional activator ToxR is a transmembrane DNA-binding protein. Cell 48:271–279 [CrossRef]
    [Google Scholar]
  39. Miller J. F., Mekalanos J. J., Falkow S. 1989; Coordinate regulation and sensory transduction in the control of bacterial virulence. Science 243:916–922 [CrossRef]
    [Google Scholar]
  40. Morris H., Schlesinger M. J, Bracha M., Yagil E. 1974; Pleiotropic effects of the mutations involved in the regulation of the Escherichia coli K-12 alkaline phosphatase. J Bacteriol 119:583–592
    [Google Scholar]
  41. Nelson E. T., Clements J. D., Finkelstein R. A. 1976; Vibrio cholerae adherence and colonization in experimental cholera: electron microscopic studies. Infect Immun 14:527–547
    [Google Scholar]
  42. Nikaido H. 1983; Proteins forming large channels from bacterial and mitochondrial outer membranes: porins and phage lambda receptor protein. Methods Enzymol 97:85–100
    [Google Scholar]
  43. Oka A., Sugisaki H., Takanami M. 1981; Nucleotide sequence of the kanamycin resistance transposon Tn903.. J Mol Biol 147:217–226 [CrossRef]
    [Google Scholar]
  44. Ostroff R. M, Wretlind B., Vasil M. L. 1989; Mutations in the hemolytic-phospholipase-C operon result in decreased virulence of Pseudomonas aeruginosa PAO1 grown under phosphate-limiting conditions. Infect Immun 57:1369–1373
    [Google Scholar]
  45. Pearson W. R., Lipman D. J. 1988; Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85:2444–2448 [CrossRef]
    [Google Scholar]
  46. Peterson K. M., Mekalanos J. J. 1988; Characterization of the Vibrio cholerae ToxR regulon – identification of novel genes involved in intestinal colonization. Infect Immun 56:2822–2829
    [Google Scholar]
  47. Raleigh E. A., Murray N. E., Revel H., Bluimenthal, R. M., Westaway D., Reith, A. D., Rigby P. W. J, Elhai J., Hanahan D. 1988; McrA and McrB restriction phenotypes of some Escherichia coli strains and implications for gene cloning. Nucleic Acids Res 16:1563 [CrossRef]
    [Google Scholar]
  48. Roy N. K., Ghosh R. K., Das J. 1982a; Monomeric alkaline phosphatase of Vibrio cholerae. J Bacteriol 150:1033–1039
    [Google Scholar]
  49. Roy N. K., Ghosh R. K., Das J. 1982b; Repression of the alkaline phosphatase of Vibrio cholerae.. J Gen Microbiol 128:349–353
    [Google Scholar]
  50. Sagar I. K., Nagesha C. N., Bhat J. V. 1981; The role of trace elements and phosphate in the synthesis of vascular-permeability factor by Vibrio cholerae.. J Med Microbiol 14:243–250 [CrossRef]
    [Google Scholar]
  51. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  52. Schnaitman C. 1974; Outer membrane proteins of Escherichia coli: evidence that the major protein of E. coli O111 outer membrane consists of four distinct polypeptide species. J Bacteriol 118:442–453
    [Google Scholar]
  53. Shine J., Dalgarno L. 1975; Determinant of cistron specificity in bacterial ribosomes. Nature 254:34–38 [CrossRef]
    [Google Scholar]
  54. Simon R., Preifer U., Püler A. 1983; A broad range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram-negative bacteria. Bio/Technology 1:784–791 [CrossRef]
    [Google Scholar]
  55. Sinai A. P., Bavoil P. M. 1993; Hyper-invasive mutants define a novel pho-regulated invasion pathway in Escherichia coli.. Mol Microbiol 10:1125–1137 [CrossRef]
    [Google Scholar]
  56. Skorupski K., Taylor R. K. 1997; Control of the ToxR virulence regulon in Vibrio cholerae by environmental stimuli. Mol Microbiol 25:1003–1011 [CrossRef]
    [Google Scholar]
  57. Stock J. B., Ninfa A. J., Stock A. M. 1989; Protein-phosphorylation and regulation of adaptive responses in bacteria. Microbiol Rev 53:450–490
    [Google Scholar]
  58. Stoker N. G., Fairweather N. F., Spratt B. G. 1982; Versatile low-copy number plasmid vector for cloning in Escherichia coli.. Gene 18:335–341 [CrossRef]
    [Google Scholar]
  59. Taylor R. K., Miller V. L., Furlong D. B., Mekalanos J. J. 1987; Use of phoA gene fusions to identify a pilus colonization factor coordinately regulated with cholera toxin. Proc Natl Acad Sci USA 84:2833–2837 [CrossRef]
    [Google Scholar]
  60. Tommassen J., Lugtenberg B. 1980; Outer membrane protein e of Escherichia coli K-12 is co-regulated with alkaline phosphatase. J Bacteriol 143:151–157
    [Google Scholar]
  61. Torriani A. 1960; Influence of inorganic phosphate (Pi) on formation of phosphatases by Escherichia coli.. Fed Proc 18:339
    [Google Scholar]
  62. VanBogelen R. A., Olson E. R., Wanner B. L., Neidhardt F. C. 1996; Global analysis of proteins synthesized during phosphorus restriction in Escherichia coli. J Bacteriol 178:4344–4366
    [Google Scholar]
  63. Vieira J., Messing J. 1982; The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic primers. Gene 19:259–268 [CrossRef]
    [Google Scholar]
  64. Wanner B. L. 1986; Novel regulatory mutants of the phosphate regulon in Escherichia coli K-12. J Mol Biol 191:39–58 [CrossRef]
    [Google Scholar]
  65. Wanner B. L. 1996; Phosphorous assimilation and control of the phosphate regulon. In Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology pp 1357–1381Edited by Neidhardt F. C. others Washington DC: American Society for Microbiology;
    [Google Scholar]
  66. Wanner B. L., McSharry R. 1982; Phosphate-controlled gene expression in Escherichia coli K12 using mudl-directed lacZ fusions. J Mol Biol 158:347–363 [CrossRef]
    [Google Scholar]
  67. Winans S. C. 1990; Transcriptional induction of an Agrobacterium regulatory gene at tandem promoters by plant-released phenolic compounds, phosphate starvation, and acidic growth media. J Bacteriol 172:2433–2438
    [Google Scholar]
  68. Wren B. W., Colby S. M., Cubberley R. R., Pallen M. J. 1992; Degenerate PCR primers for the amplification of fragments from genes encoding response regulators from a range of pathogenic bacteria. FEMS Microbiol Lett 99:287–291 [CrossRef]
    [Google Scholar]
  69. Yagil E. M, Bracha M., Lifshitz Y. 1975; The regulatory nature of the phoB gene for alkaline phosphatase synthesis in Escherichia coli.. Mol Gen Genet 137:11–16
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-145-9-2463
Loading
/content/journal/micro/10.1099/00221287-145-9-2463
Loading

Data & Media loading...

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