1887

Abstract

Computer searches were carried out of the gonococcal and meningococcal genome databases for previously unknown members of the onB-ependent amily (Tdf) of outer-membrane receptor proteins. Seven putative non-contiguous genes were found and three of these (identified in gonococcal strain FA1090) were chosen for further study. Consensus motif analysis of the peptide sequences was consistent with the three genes encoding TonB-dependent receptors. In view of the five previously characterized TonB-dependent proteins of pathogenic neisseriae, the putative genes were labelled , and . TdfF had homology with the siderophore receptors FpvA of and FhuE of , whereas TdfG and TdfH had homology with the haemophore receptor HasR of . The aim of this project was to characterize these proteins and determine their expression, regulation, distribution and surface exposure. Strain surveys of iron-stressed commensal and pathogenic neisseriae revealed that TdfF is unlikely to be expressed, TdfG is expressed by gonococci only and that TdfH is expressed by both meningococci and gonococci. Expression of TdfH was unaffected by iron availability. Susceptibility of TdfH to cleavage by proteases in live gonococci was consistent with surface exposure of this protein. TdfH may function as a TonB-dependent receptor for a non-iron nutrient source. Furthermore, TdfH is worthy of future investigation as a potential meningococcal vaccine candidate as it is a highly conserved, widely distributed and surface-exposed outer-membrane protein.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-147-5-1277
2001-05-01
2020-08-08
Loading full text...

Full text loading...

/deliver/fulltext/micro/147/5/1471277a.html?itemId=/content/journal/micro/10.1099/00221287-147-5-1277&mimeType=html&fmt=ahah

References

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. 1990; Basic local alignment search tool. J Mol Biol215:403–410[CrossRef]
    [Google Scholar]
  2. Beucher M., Sparling P. F. 1995; Cloning, sequencing and characterization of the gene encoding FrpB, a major iron-regulated, outer membrane protein of Neisseria gonorrhoeae . J Bacteriol177:2041–2049
    [Google Scholar]
  3. Bitter W., Marugg J. D., Tommassen J., Weisbeek P. J, de, Weger L. A.. 1991; The ferric-pseudobactin receptor PupA of Pseudomonas putida WCS358: homology to TonB-dependent Escherichia coli receptors and specificity of the protein. Mol Microbiol5:647–655[CrossRef]
    [Google Scholar]
  4. Braun V. 1995; Energy-coupled transport and signal transduction through the Gram-negative outer membrane via TonB-ExbB-ExbD-dependent receptor proteins. FEMS Microbiol Rev16:295–307[CrossRef]
    [Google Scholar]
  5. Braun V., Killmann H. 1999; Bacterial solutions to the iron-supply problem. Trends Biochem Sci24:104–109[CrossRef]
    [Google Scholar]
  6. Carson S. D., Klebba P. E., Newton S. M., Sparling P. F. 1999; Ferric enterobactin binding and utilization by Neisseria gonorrhoeae . J Bacteriol181:2895–2901
    [Google Scholar]
  7. Chen C. J., Sparling P. F., Lewis L. A., Dyer D. W., Elkins C. 1996; Identification and purification of a hemoglobin-binding outer membrane protein from Neisseria gonorrhoeae . Infect Immun64:5008–5014
    [Google Scholar]
  8. Chen C. Y., Morse S. A. 2000; Identification and characterization of a high-affinity zinc uptake system in Neisseria gonorrhoeae . In Neisseria 2000; Twelfth International Pathogenic Neisseria Conference Edited by Rice P., Sparling P. F., Shafer W.. Galveston, TX: Health Care Communications;
    [Google Scholar]
  9. Cornelissen C. N., Sparling P. F. 1996; Binding and surface exposure characteristics of the gonococcal transferrin receptor are dependent on both transferrin-binding proteins. J Bacteriol178:1437–1444
    [Google Scholar]
  10. Cornelissen C. N., Biswas G. D., Tsai J., Paruchuri D. K., Thompson S. A., Sparling P. F. 1992; Gonococcal transferrin-binding protein 1 is required for transferrin utilization and is homologous to TonB-dependent outer membrane receptors. J Bacteriol174:5788–5797
    [Google Scholar]
  11. Correia F. F., Inouye S., Inouye M. 1986; A 26-base-pair repetitive sequence specific for Neisseria gonorrhoeae and Neisseria meningitidis genomic DNA. J Bacteriol167:1009–1015
    [Google Scholar]
  12. Correia F. F., Inouye S., Inouye M. 1988; A family of small repeated elements with some transposon-like properties in the genome of Neisseria gonorrhoeae . J Biol Chem263:12194–12198
    [Google Scholar]
  13. Dempsey J. F., Litaker W., Madhure A., Snodgrass T. L., Cannon J. G. 1991; Physical map of the chromosome of Neisseria gonorrhoeae FA1090 with locations of genetic markers, including opa and pil genes. J Bacteriol173:5476–5486
    [Google Scholar]
  14. Dyer D. W., McKenna W., Woods J. P., Sparling P. F. 1987; Isolation by streptonigrin enrichment and characterization of a transferrin-specific iron uptake mutant of Neisseria meningitidis . Microb Pathog3:351–363[CrossRef]
    [Google Scholar]
  15. Fiss E. H., Stanley-Samuelson P., Neilands J. B. 1982; Properties and proteolysis of ferric enterobactin outer membrane receptor in Escherichia coli K12. Biochemistry21:4517–4522[CrossRef]
    [Google Scholar]
  16. Fleischmann R. D., Adams M. D., White O.. 37 other authors 1995; Whole-genome random sequencing and assembly of Haemophilus influenzae Rd. Science269:496–512[CrossRef]
    [Google Scholar]
  17. Griffiss J. M., Yamasaki R., Estabrook M., Kim J. J. 1991; Meningococcal molecular mimicry and the search for an ideal vaccine. Trans R Soc Trop Med Hyg85 Suppl: 132–36
    [Google Scholar]
  18. Guerinot M. L. 1994; Microbial iron transport. Annu Rev Microbiol48:743–772[CrossRef]
    [Google Scholar]
  19. Heinrichs D. E., Poole K. 1993; Cloning and sequence analysis of a gene ( pchR ) encoding an AraC family activator of pyochelin and ferripyochelin receptor synthesis in Pseudomonas aeruginosa . J Bacteriol175:5882–5889
    [Google Scholar]
  20. Heinrichs D. E., Poole K. 1996; PchR, a regulator of ferripyochelin receptor gene ( fptA ) expression in Pseudomonas aeruginosa , functions both as an activator and as a repressor. J Bacteriol178:2586–2592
    [Google Scholar]
  21. Heller J. K., Kadner R. J., Gunter K. 1988; Supression of btuB 451 mutation by mutations in the tonB gene suggests a direct interaction between TonB and TonB-dependent receptor proteins in the outer membrane of Escherichia coli . Gene64:147–153[CrossRef]
    [Google Scholar]
  22. Heller K., Kadner R. J. 1985; Nucleotide sequence of the gene for the vitamin B12 receptor protein in the outer membrane of Escherichia coli . J Bacteriol161:904–908
    [Google Scholar]
  23. Hobbs M. M., Alcorn T. M., Davis R. H., Fischer W., Thomas J. C., Martin I., Ison C., Sparling P. F., Cohen M. S. 1999; Molecular typing of Neisseria gonorrhoeae causing repeated infections: evolution of porin during passage within a community. J Infect Dis179:371–381[CrossRef]
    [Google Scholar]
  24. Kadner R. J. 1990; Vitamin B12 transport in Escherichia coli : energy coupling between membranes. Mol Microbiol4:2027–2033[CrossRef]
    [Google Scholar]
  25. Kammen H. O., Marvel C. C., Hardy L., Penhoet E. E. 1988; Purification, structure, and properties of Escherichia coli tRNA pseudouridine synthase I. J Biol Chem263:2255–2263
    [Google Scholar]
  26. Kellogg D. S. Jr, Peacock W. L. Jr, Deacon W. E., Brown L., Pirkle C. I. 1963; Neisseria gonorrhoeae . I. Virulence genetically linked to clonal variation. J Bacteriol85:1274–1279
    [Google Scholar]
  27. Klebba P. E., Rutz J. M., Liu J., Murphy C. K. 1993; Mechanisms of TonB-catalyzed iron transport through the enteric bacterial cell envelope. J Bioenerg Biomembr25:603–611
    [Google Scholar]
  28. Klee S. R., Nassif X., Kusecek B., Merker P., Beretti J. L., Achtman M., Tinsley C. R. 2000; Molecular and biological analysis of eight genetic islands that distinguish Neisseria meningitidis from the closely related pathogen Neisseria gonorrhoeae . Infect Immun68:2082–2095[CrossRef]
    [Google Scholar]
  29. Lee B. C. 1992; Isolation of haemin-binding proteins of Neisseria gonorrhoeae . J Med Microbiol36:121–127[CrossRef]
    [Google Scholar]
  30. Lee B. C. 1994; Isolation and characterization of the haemin-binding proteins from Neisseria meningitidis . Microbiology140:1473–1480[CrossRef]
    [Google Scholar]
  31. Lee B. C., Levesque S. 1997; A monoclonal antibody directed against the 97-kilodalton gonococcal hemin-binding protein inhibits hemin utilization by Neisseria gonorrhoeae . Infect Immun65:2970–2974
    [Google Scholar]
  32. Letoffe S., Ghigo J. M., Wandersman C. 1994; Iron acquisition from heme and hemoglobin by Serratia marcescens extracellular protein. Proc Natl Acad Sci USA91:9876–9880[CrossRef]
    [Google Scholar]
  33. Lewis D. A., Klesney-Tait J., Lumbley S. R., Ward C. K., Latimer J. L., Ison C. A., Hansen E. J. 1999; Identification of the znuA -encoded periplasmic zinc transport protein of Haemophilus ducreyi . Infect Immun67:5060–5068
    [Google Scholar]
  34. Lewis L. A., Dyer D. D. 1995; Identification of an iron-regulated outer membrane protein of Neisseria meningitidis involved in the utilization of hemoglobin complexed to haptoglobin. J Bacteriol177:1299–1306
    [Google Scholar]
  35. Li C., Clarke S. 1992; A protein methyltransferase specific for altered aspartyl residues is important in Escherichia coli stationary-phase survival and heat-shock resistance. Proc Natl Acad Sci U S A89:9885–9889[CrossRef]
    [Google Scholar]
  36. Ling M., Allen S. W., Wood J. M. 1994; Sequence analysis identifies the proline dehydrogenase and delta-1-pyrroline-5-carboxylate dehydrogenase domains of the multifunctional Escherichia coli PutA protein. J Mol Biol243:950–956[CrossRef]
    [Google Scholar]
  37. Litwin C. M., Calderwood S. B. 1993; Role of iron in regulation of virulence genes. Clin Microbiol Rev6:137–149
    [Google Scholar]
  38. Lundrigan M. D., Kadner R. J. 1986; Nucleotide sequence of the gene for the ferrienterochelin receptor FepA in Escherichia coli . Homology among outer membrane receptors that interact with TonB. J Biol Chem261:10797–10801
    [Google Scholar]
  39. Meyer T. F., Mlawer N., So M. 1982; Pilus expression in Neisseria gonorrhoeae involves chromosomal rearrangement. Cell30:45–52[CrossRef]
    [Google Scholar]
  40. Moeck G. S., Coulton J. W. 1998; TonB-dependent iron acquisition: mechanisms of siderophore-mediated active transport. Mol Microbiol28:675–681
    [Google Scholar]
  41. Nakano Y., Yoshida Y., Yamshita Y., Doga T. 1995; Construction of a series of pACYC-derived plasmid vectors. Gene162:157–158[CrossRef]
    [Google Scholar]
  42. Parkhill J., Achtman M., James K. D.. 25 other authors 2000; Complete DNA sequence of a serogroup A strain of Neisseria meningitidis Z2491. Nature404:502–506[CrossRef]
    [Google Scholar]
  43. Patzer S. I., Hantke K. 1998; The ZnuABC high-affinity zinc uptake system and its regulator Zur in Escherichia coli . Mol Microbiol28:1199–1210[CrossRef]
    [Google Scholar]
  44. Payne S. M. 1988; Iron and virulence in the family Enterobacteriaceae . Crit Rev Microbiol16:81–111[CrossRef]
    [Google Scholar]
  45. Pizza M., Scarlato V., Masignani V.. 33 other authors 2000; Identification of vaccine candidates against serogroup B meningococcus by whole-genome sequencing. Science287:1816–1820[CrossRef]
    [Google Scholar]
  46. Poole K., Neshat S., Krebes K., Heinrichs D. E. 1993; Cloning and nucleotide sequence analysis of the ferripyoverdine receptor gene fpvA of Pseudomonas aeruginosa . J Bacteriol175:4597–4604
    [Google Scholar]
  47. Postle K. 1999; Active transport by customized beta-barrels. Nat Struct Biol6:3–6[CrossRef]
    [Google Scholar]
  48. Prentki P., Kirsch H. M. 1984; In vitro insertional mutagenesis with a selectable DNA fragment. Gene29:303–313[CrossRef]
    [Google Scholar]
  49. Pressler U., Staudenmaier H., Zimmermann L., Braun V. 1988; Genetics of the iron dicitrate transport system of Escherichia coli . J Bacteriol170:2716–2724
    [Google Scholar]
  50. Prince A., Wood M. S., Cacalano G. S., Chin N. X. 1988; Isolation and characterization of a penicillinase from Pseudomonas cepacia 249. Antimicrob Agents Chemother32:838–843[CrossRef]
    [Google Scholar]
  51. Qi H. L., Tai J. Y., Blake M. S. 1994; Expression of large amounts of neisserial porin proteins in Escherichia coli and refolding of the proteins into native trimers. Infect Immun62:2432–2439
    [Google Scholar]
  52. Richardson A. R., Stojiljkovic I. 1999; HmbR, a hemoglobin-binding outer membrane protein of Neisseria meningitidis , undergoes phase variation. J Bacteriol181:2067–2074
    [Google Scholar]
  53. Richardson P. T., Park S. F. 1995; Enterochelin acquisition in Campylobacter coli : characterization of components of a binding-protein-dependent transport system. Microbiology141:3181–3191[CrossRef]
    [Google Scholar]
  54. Sauer M., Hantke K., Braun V. 1990; Sequence of the fhuE outer-membrane receptor gene of Escherichia coli K12 and properties of mutants. Mol Microbiol4:427–437[CrossRef]
    [Google Scholar]
  55. Saunders N. J., Jeffries A. C., Peden J. F., Hood D. W., Tettelin H., Rappuoli R., Moxon E. R. 2000; Repeat-associated phase variable genes in the complete genome sequence of Neisseria meningitidis strain MC58. Mol Microbiol37:207–215[CrossRef]
    [Google Scholar]
  56. Schryvers A. B., Morris L. J. 1988a; Identification and characterization of the transferrin receptor from Neisseria meningitidis . Mol Microbiol2:281–288[CrossRef]
    [Google Scholar]
  57. Schryvers A. B., Morris L. J. 1988b; Identification and characterization of the human lactoferrin-binding protein from Neisseria meningitidis . Infect Immun56:1144–1149
    [Google Scholar]
  58. Schryvers A. B., Stojiljkovic I. 1999; Iron acquisition systems in the pathogenic Neisseria. Mol Microbiol32:1117–1123[CrossRef]
    [Google Scholar]
  59. Schultz J. E., Matin A. 1991; Molecular and functional characterization of a carbon starvation gene of Escherichia coli . J Mol Biol218:129–140[CrossRef]
    [Google Scholar]
  60. Stojiljkovic I., Hantke K. 1992; Haemin uptake system of Yersinia enterocolitica : similarities with other TonB-dependent systems in Gram-negative bacteria. EMBO J11:4359–4367
    [Google Scholar]
  61. Stojiljkovic I., Hwa V., de Saint Martin L., O’Gaora P., Nassif X., Heffron F., So M. 1995; The Neisseria meningitidis haemoglobin receptor: its role in iron utilization and virulence. Mol Microbiol15:531–541[CrossRef]
    [Google Scholar]
  62. Stojiljkovic I., Larson J., Hwa V., Anic S., So M. 1996; HmbR outer membrane receptors of pathogenic Neisseria spp.: iron-regulated, hemoglobin-binding proteins with a high level of primary structure conservation. J Bacteriol178:4670–4678
    [Google Scholar]
  63. Struyve M., Moons M., Tommassen J. 1991; Carboxyl-terminal phenylalanine is essential for the correct assembly of a bacterial outer membrane protein. J Mol Biol218:141–148[CrossRef]
    [Google Scholar]
  64. Suhr M., Benz I., Schmidt M. A. 1996; Processing of the AIDA-I precursor: removal of AIDAc and evidence for the outer membrane anchoring as a beta-barrel structure. Mol Microbiol22:31–42[CrossRef]
    [Google Scholar]
  65. Tettelin H., Saunders N. J., Heidelberg J.. 39 other authors 2000; Complete genome sequence of Neisseria meningitidis serogroup B strain MC58. Science287:1809–1815[CrossRef]
    [Google Scholar]
  66. Thomas C. E., Sparling P. F. 1996; Isolation and analysis of a fur mutant of Neisseria gonorrhoeae . J Bacteriol178:4224–4232
    [Google Scholar]
  67. Tinoco I. Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. 1973; Improved estimation of secondary structure in ribonucleic acids. Nat New Biol246:40–41[CrossRef]
    [Google Scholar]
  68. Towbin J., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedures and some applications. Proc Natl Acad Sci USA76:4350–4354[CrossRef]
    [Google Scholar]
  69. Turner P. C., Thomas C. E., Elkins C., Clary S., Sparling P. F. 1998; Neisseria gonorrhoeae heme biosynthetic mutants utilize heme and hemoglobin as a heme source but fail to grow within epithelial cells. Infect Immun66:5215–5223
    [Google Scholar]
  70. West S. E., Sparling P. F. 1985; Response of Neisseria gonorrhoeae to iron limitation: alterations in expression of membrane proteins without apparent siderophore production. Infect Immun477:388–394
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-147-5-1277
Loading
/content/journal/micro/10.1099/00221287-147-5-1277
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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