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J Bacteriol. 1996 November; 178(22): 6555–6563.
PMCID: PMC178542

BfpB, an outer membrane lipoprotein required for the biogenesis of bundle-forming pili in enteropathogenic Escherichia coli.

Abstract

The bundle-forming pili (BFP) of enteropathogenic Escherichia coli are believed to play a role in pathogenesis by causing the formation of bacterial microcolonies that bind epithelial surfaces of the small intestine. This in vivo process is mimicked in vitro by the autoaggregation and localized adherence phenotypes. Expression of BFP, a member of the type IV pilus family, requires the enteroadherence factor (EAF) plasmid, which contains bfpA, the gene that encodes the principal structural subunit of BFP. Immediately downstream of bfpA are 13 open reading frames transcribed in the same direction as bfpA; together with bfpA, these compose the bfp gene cluster. Disruption of bfpB, the second open reading frame downstream of bfpA, was performed by allelic exchange. The resulting mutant, B171-8deltaB, did not exhibit the autoaggregation or localized adherence phenotype or produce BFP filaments. Thus, BfpB is required for pilus biogenesis. However, BfpA was produced at wild-type levels and processed normally by B171-8deltaB, indicating that BfpB acts at a step in the BFP biogenic pathway after production and processing of the structural subunit. Biochemical and cell fractionation studies showed that BfpB is a 58-kDa lipoprotein that is located primarily in the outer membrane. Assays of bfpA and bfpB mRNAs and protein expression showed that both genes are cotranscribed as part of an environmentally responsive operon that is regulated by growth phase and ammonium.

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Selected References

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  • Aiba H, Adhya S, de Crombrugghe B. Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem. 1981 Nov 25;256(22):11905–11910. [PubMed]
  • Baldini MM, Kaper JB, Levine MM, Candy DC, Moon HW. Plasmid-mediated adhesion in enteropathogenic Escherichia coli. J Pediatr Gastroenterol Nutr. 1983;2(3):534–538. [PubMed]
  • Boeke JD, Model P. A prokaryotic membrane anchor sequence: carboxyl terminus of bacteriophage f1 gene III protein retains it in the membrane. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5200–5204. [PubMed]
  • Brown RC, Taylor RK. Organization of tcp, acf, and toxT genes within a ToxT-dependent operon. Mol Microbiol. 1995 May;16(3):425–439. [PubMed]
  • Chou PY, Fasman GD. Conformational parameters for amino acids in helical, beta-sheet, and random coil regions calculated from proteins. Biochemistry. 1974 Jan 15;13(2):211–222. [PubMed]
  • Echeverria P, Taylor DN, Donohue-Rolfe A, Supawat K, Ratchtrachenchai O, Kaper J, Keusch GT. HeLa cell adherence and cytotoxin production by enteropathogenic Escherichia coli isolated from infants with diarrhea in Thailand. J Clin Microbiol. 1987 Aug;25(8):1519–1523. [PMC free article] [PubMed]
  • Girón JA, Ho AS, Schoolnik GK. An inducible bundle-forming pilus of enteropathogenic Escherichia coli. Science. 1991 Nov 1;254(5032):710–713. [PubMed]
  • Gomes TA, Vieira MA, Wachsmuth IK, Blake PA, Trabulsi LR. Serotype-specific prevalence of Escherichia coli strains with EPEC adherence factor genes in infants with and without diarrhea in São Paulo, Brazil. J Infect Dis. 1989 Jul;160(1):131–135. [PubMed]
  • Gomes TA, Blake PA, Trabulsi LR. Prevalence of Escherichia coli strains with localized, diffuse, and aggregative adherence to HeLa cells in infants with diarrhea and matched controls. J Clin Microbiol. 1989 Feb;27(2):266–269. [PMC free article] [PubMed]
  • Hardie KR, Lory S, Pugsley AP. Insertion of an outer membrane protein in Escherichia coli requires a chaperone-like protein. EMBO J. 1996 Mar 1;15(5):978–988. [PubMed]
  • Hobbs M, Collie ES, Free PD, Livingston SP, Mattick JS. PilS and PilR, a two-component transcriptional regulatory system controlling expression of type 4 fimbriae in Pseudomonas aeruginosa. Mol Microbiol. 1993 Mar;7(5):669–682. [PubMed]
  • Hobbs M, Mattick JS. Common components in the assembly of type 4 fimbriae, DNA transfer systems, filamentous phage and protein-secretion apparatus: a general system for the formation of surface-associated protein complexes. Mol Microbiol. 1993 Oct;10(2):233–243. [PubMed]
  • Hu NT, Hung MN, Liao CT, Lin MH. Subcellular location of XpsD, a protein required for extracellular protein secretion by Xanthomonas campestris pv. campestris. Microbiology. 1995 Jun;141(Pt 6):1395–1406. [PubMed]
  • Kaufman MR, Shaw CE, Jones ID, Taylor RK. Biogenesis and regulation of the Vibrio cholerae toxin-coregulated pilus: analogies to other virulence factor secretory systems. Gene. 1993 Apr 15;126(1):43–49. [PubMed]
  • Kazmierczak BI, Mielke DL, Russel M, Model P. pIV, a filamentous phage protein that mediates phage export across the bacterial cell envelope, forms a multimer. J Mol Biol. 1994 Apr 29;238(2):187–198. [PubMed]
  • Knutton S, Baldini MM, Kaper JB, McNeish AS. Role of plasmid-encoded adherence factors in adhesion of enteropathogenic Escherichia coli to HEp-2 cells. Infect Immun. 1987 Jan;55(1):78–85. [PMC free article] [PubMed]
  • Knutton S, Baldwin T, Williams PH, McNeish AS. Actin accumulation at sites of bacterial adhesion to tissue culture cells: basis of a new diagnostic test for enteropathogenic and enterohemorrhagic Escherichia coli. Infect Immun. 1989 Apr;57(4):1290–1298. [PMC free article] [PubMed]
  • Knutton S, Lloyd DR, McNeish AS. Adhesion of enteropathogenic Escherichia coli to human intestinal enterocytes and cultured human intestinal mucosa. Infect Immun. 1987 Jan;55(1):69–77. [PMC free article] [PubMed]
  • Levine MM, Nataro JP, Karch H, Baldini MM, Kaper JB, Black RE, Clements ML, O'Brien AD. The diarrheal response of humans to some classic serotypes of enteropathogenic Escherichia coli is dependent on a plasmid encoding an enteroadhesiveness factor. J Infect Dis. 1985 Sep;152(3):550–559. [PubMed]
  • Linderoth NA, Model P, Russel M. Essential role of a sodium dodecyl sulfate-resistant protein IV multimer in assembly-export of filamentous phage. J Bacteriol. 1996 Apr;178(7):1962–1970. [PMC free article] [PubMed]
  • McConnell MM, Chart H, Scotland SM, Smith HR, Willshaw GA, Rowe B. Properties of adherence factor plasmids of enteropathogenic Escherichia coli and the effect of host strain on expression of adherence to HEp-2 cells. J Gen Microbiol. 1989 May;135(5):1123–1134. [PubMed]
  • Miller VL, Mekalanos JJ. 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. 1988 Jun;170(6):2575–2583. [PMC free article] [PubMed]
  • Nataro JP, Baldini MM, Kaper JB, Black RE, Bravo N, Levine MM. Detection of an adherence factor of enteropathogenic Escherichia coli with a DNA probe. J Infect Dis. 1985 Sep;152(3):560–565. [PubMed]
  • Nataro JP, Maher KO, Mackie P, Kaper JB. Characterization of plasmids encoding the adherence factor of enteropathogenic Escherichia coli. Infect Immun. 1987 Oct;55(10):2370–2377. [PMC free article] [PubMed]
  • Nataro JP, Scaletsky IC, Kaper JB, Levine MM, Trabulsi LR. Plasmid-mediated factors conferring diffuse and localized adherence of enteropathogenic Escherichia coli. Infect Immun. 1985 May;48(2):378–383. [PMC free article] [PubMed]
  • Ogierman MA, Zabihi S, Mourtzios L, Manning PA. Genetic organization and sequence of the promoter-distal region of the tcp gene cluster of Vibrio cholerae. Gene. 1993 Apr 15;126(1):51–60. [PubMed]
  • Osborn MJ, Munson R. Separation of the inner (cytoplasmic) and outer membranes of Gram-negative bacteria. Methods Enzymol. 1974;31:642–653. [PubMed]
  • Paulozzi LJ, Johnson KE, Kamahele LM, Clausen CR, Riley LW, Helgerson SD. Diarrhea associated with adherent enteropathogenic Escherichia coli in an infant and toddler center, Seattle, Washington. Pediatrics. 1986 Mar;77(3):296–300. [PubMed]
  • Polotsky YE, Dragunskaya EM, Seliverstova VG, Avdeeva TA, Chakhutinskaya MG, Kétyi I, Vertényl A, Ralovich B, Emödy L, Málovics I, et al. Pathogenic effect of enterotoxigenic Escherichia coli and Escherichia coli causing infantile diarrhoea. Acta Microbiol Acad Sci Hung. 1977;24(3):221–236. [PubMed]
  • Puente JL, Bieber D, Ramer SW, Murray W, Schoolnik GK. The bundle-forming pili of enteropathogenic Escherichia coli: transcriptional regulation by environmental signals. Mol Microbiol. 1996 Apr;20(1):87–100. [PubMed]
  • Pugsley AP. The complete general secretory pathway in gram-negative bacteria. Microbiol Rev. 1993 Mar;57(1):50–108. [PMC free article] [PubMed]
  • Quandt J, Hynes MF. Versatile suicide vectors which allow direct selection for gene replacement in gram-negative bacteria. Gene. 1993 May 15;127(1):15–21. [PubMed]
  • Rothbaum R, McAdams AJ, Giannella R, Partin JC. A clinicopathologic study of enterocyte-adherent Escherichia coli: a cause of protracted diarrhea in infants. Gastroenterology. 1982 Aug;83(2):441–454. [PubMed]
  • Rothbaum RJ, Partin JC, Saalfield K, McAdams AJ. An ultrastructural study of enteropathogenic Escherichia coli infection in human infants. Ultrastruct Pathol. 1983 Jun;4(4):291–304. [PubMed]
  • Russel M, Kaźmierczak B. Analysis of the structure and subcellular location of filamentous phage pIV. J Bacteriol. 1993 Jul;175(13):3998–4007. [PMC free article] [PubMed]
  • Sohel I, Puente JL, Murray WJ, Vuopio-Varkila J, Schoolnik GK. Cloning and characterization of the bundle-forming pilin gene of enteropathogenic Escherichia coli and its distribution in Salmonella serotypes. Mol Microbiol. 1993 Feb;7(4):563–575. [PubMed]
  • Sohel I, Puente JL, Ramer SW, Bieber D, Wu CY, Schoolnik GK. Enteropathogenic Escherichia coli: identification of a gene cluster coding for bundle-forming pilus morphogenesis. J Bacteriol. 1996 May;178(9):2613–2628. [PMC free article] [PubMed]
  • Stone KD, Zhang HZ, Carlson LK, Donnenberg MS. A cluster of fourteen genes from enteropathogenic Escherichia coli is sufficient for the biogenesis of a type IV pilus. Mol Microbiol. 1996 Apr;20(2):325–337. [PubMed]
  • Strom MS, Lory S. Structure-function and biogenesis of the type IV pili. Annu Rev Microbiol. 1993;47:565–596. [PubMed]
  • Ulshen MH, Rollo JL. Pathogenesis of escherichia coli gastroenteritis in man--another mechanism. N Engl J Med. 1980 Jan 10;302(2):99–101. [PubMed]
  • Vuopio-Varkila J, Schoolnik GK. Localized adherence by enteropathogenic Escherichia coli is an inducible phenotype associated with the expression of new outer membrane proteins. J Exp Med. 1991 Nov 1;174(5):1167–1177. [PMC free article] [PubMed]
  • Wang RF, Kushner SR. Construction of versatile low-copy-number vectors for cloning, sequencing and gene expression in Escherichia coli. Gene. 1991 Apr;100:195–199. [PubMed]

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