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The DraC usher in Dr fimbriae biogenesis of uropathogenic E. coli Dr+ strains

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Abstract

Biogenesis of Dr fimbriae encoded by the dra gene cluster of uropathogenic Escherichia coli strains requires the chaperone-usher pathway. This secretion system is based on two non-structural assembly components, the DraB periplasmic chaperone and DraC outer-membrane usher. The DraB controls the folding of DraE subunits, and DraC forms the assembly and secretion platform for polymerization of subunits in linear fibers. In this study, mutagenesis of the DraC N-terminus was undertaken to select residues critical for Dr fimbriae bioassembly. The DraC-F4A, DraC-C64, DraC-C100A and DraC-W142A significantly reduced the adhesive ability of E. coli strains. The biological activity of the DraC mutants as a assembly platform for Dr fimbriae polymerization was verified by agglutination of human erythrocytes and adhesion to DAF localized at the surface of CHO-DAF+ and HeLa cells. The residue F4 of the DraC usher conserved among FGL and FGS chaperone-assembled adhesive organelles can be used to design pillicides blocking the biogenesis of Dr fimbriae. Because the draC and afaC-III genes share 100% identity the range of the virulence determinant inhibitors could also be extended to E. coli strains encoding afa-3 gene cluster. The investigations performed showed that the usher N-terminus plays an important role in biogenesis of complete fiber.

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References

  • Aberg V, Almqvist F (2007) Pilicides—small molecules targeting bacterial virulence. Org Biomol Chem 5:1827–1834

    Article  PubMed  CAS  Google Scholar 

  • Berger CN, Billker O, Meyer TF, Servin AL, Kansau I (2004) Differential recognition of the carcinoembryonic antigen family by Afa/Dr adhesins of diffusely adhering Echerichia coli (Afa/Dr DAEC). Mol Microbiol 52:963–983

    Article  CAS  PubMed  Google Scholar 

  • Capitani G, Eidam O, Glockshuber R, Grütter MG (2006a) Structural and functional insights into the assembly of type 1 pili from Escherichia coli. Microbes Infect 8:2284–2290

    Article  CAS  PubMed  Google Scholar 

  • Capitani G, Eidam O, Grütter MG (2006b) Evidence for a novel domain of bacterial outer membrane ushers. Proteins 65:816–823

    Article  CAS  PubMed  Google Scholar 

  • Carnoy C, Moseley SL (1997) Mutational analysis of receptor binding mediated by the Dr family of Escherichia coli adhesins. Mol Microbiol 23:365–379

    Article  CAS  PubMed  Google Scholar 

  • Garcia MI, Labigne A, LeBouguenec C (1994) Nucleotide sequence of the afimbrial-adhesin-encoding afa-3 gene cluster and its translocation via flankig IS1 insertion sequences. J Bacteriol 176:7601–7613

    CAS  PubMed  Google Scholar 

  • Goluszko P, Selvarangan R, Popov V, Pham T, Wen JW, Singhal J (1999) Decay-accelerating factor and cytoskeleton redistribution pattern in HeLa cells infected with recombinant Escherichia coli strains expressing Dr family of adhesins. Infect Immun 67:3989–3997

    CAS  PubMed  Google Scholar 

  • Guignot J, Hudault S, Kansau I, Chau I, Servin AL (2009) DAF and CEACAMs receptor-mediated internalization and lifestyle of Afa/Dr diffusely adhering Escherichia coli into epithelial cells. Infect Immun 77:517–531

    Article  CAS  PubMed  Google Scholar 

  • Henderson N, So SS, Martin C, Kulkarni R, Thanassi D (2004) Topology of the outer membrane usher PapC determined by site-directed fluorescence labeling. J Biol Chem 51:53747–53754

    Article  CAS  Google Scholar 

  • Jędrzejczak R, Dauter Z, Dauter M, Piątek R, Zalewska B, Mróz M, Bury K, Nowicki BJ, Kur J (2006) Structure of DraD invasin from uropathogenic Escherichia coli: a dimer with swapped beta-tails. Acta Crystallogr D Biol Crystallogr 62:157–164

    Article  PubMed  CAS  Google Scholar 

  • Johnson JR (1991) Virulence factors in Escherichia coli urinary tract infection. Clin Microbiol Rev 4:80–128

    CAS  PubMed  Google Scholar 

  • Korotkova N, Yang Y, Le Trong I, Cota E, Demeler B, Marchant J, Thomas WE, Stenkamp RE, Moseley SL, Matthews S (2008a) Binding of Dr adhesins of Escherichia coli to carcinoembryonic antigen triggers receptor disssociation. Mol Microbiol 67:420–434

    Article  CAS  PubMed  Google Scholar 

  • Korotkova N, Yarova-Yarovaya Y, Tchesnokova V, Yazvenko N, Carl MA, Stapleton AE, Moseley SL (2008b) Escherichia coli DraE adhesin-associated bacterial internalization by epithelial cells is promoted independently by decay-accelerating factor and carcinoembryonic antigen-related cell adhesion molecule binding and does not require the DraD invasin. Infect Immun 76:3869–3880

    Article  CAS  PubMed  Google Scholar 

  • Kur M, Piatek R, Kur J (2007) A two-plasmid Escherichia coli system for expression of Dr adhesins. Protein Expr Purif 55:361–367

    Article  CAS  PubMed  Google Scholar 

  • Lee YM, Almqvist F, Hultgren SJ (2003) Targeting virulence for antimicrobial chemotherapy. Curr Opin Pharmacol 3:513–519

    Article  CAS  PubMed  Google Scholar 

  • Lublin DM, Coyne KE (1991) Phospholipid-anchored and transmembrane versions of either decay-accelerating factor or membrane cofactor protein show equal efficiency in protection from complement-mediated cell damage. J Exp Med 174:35–44

    Article  CAS  PubMed  Google Scholar 

  • Ng TW, Akman L, Osisami M, Thanassi DG (2004) The usher N terminus is the initial targeting site for chaperone-subunit complexes and participates in subsequent pilus biogenesis events. J Bacteriol 186:5321–5331

    Article  CAS  PubMed  Google Scholar 

  • Nishiyama M, Horst R, Eidam O, Herrmann T, Ignatov O, Vetsch M, Bettendorff P, Jelesarov I, Grütter MG, Wüthrich K, Glockshuber R, Capitani G (2005) Structural basis of chaperone-subunit complex recognition by the type 1 pilus assembly platform FimD. EMBO J 24:2075–2086

    Article  CAS  PubMed  Google Scholar 

  • Nowicki B, Barrish JP, Korhonen T, Hull RA, Hull SI (1987) Molecular cloning of the Escherichia coli O75X adhesin. Infect Immun 553:3168–3173

    Google Scholar 

  • Nowicki B, Moulds J, Hull R, Hull S (1988) A hemagglutinin of uropathogenic Escherichia coli recognizes the Dr blood group antigen. Infect Immun 56:1057–1060

    CAS  PubMed  Google Scholar 

  • Nowicki B, Svanborg-Eden C, Hull R, Hull S (1989) Molecular analysis and epidemiology of the Dr hemagglutinin of uropathogenic Escherichia coli. Infect Immun 57:446–451

    CAS  PubMed  Google Scholar 

  • Nowicki B, Martens M, Hart A, Nowicki S (1994) Gestational age-dependent distribution of Escherichia coli fimbriae in pregnant patients with pyelonephritis. Ann N Y Acad Sci 730:290–291

    Article  CAS  PubMed  Google Scholar 

  • Nowicki B, Selvarangan R, Nowicki S (2001) Family of Escherichia coli Dr adhesins: decay-accelerating factor receptor recognition and invasiveness. J Infect Dis 183 (Suppl. 1):S24–S27

    Article  Google Scholar 

  • Pham TQ, Goluszko P, Popov V, Nowicki S, Nowicki BJ (1997) Molecular cloning and characterization of Dr-II, a nonfimbrial adhesin-I-like adhesin isolated from gestational pyelonephritis-associated Escherichia coli that binds to decay-accelerating factor. Infect Immun 65:4309–4318

    CAS  PubMed  Google Scholar 

  • Piątek R, Zalewska B, Kolaj O, Ferens M, Nowicki B, Kur J (2005a) Molecular aspects of biogenesis of Escherichia coli Dr Fimbriae: characterization of DraB-DraE complexes. Infect Immun 73:135–145

    Article  PubMed  CAS  Google Scholar 

  • Piątek R, Zalewska B, Bury K, Kur J (2005b) The chaperone-usher pathway of bacterial adhesin biogenesis—from molecular mechanism to strategies of anti-bacterial prevention and modern vaccine design. Acta Biochim Pol 52:639–646

    PubMed  Google Scholar 

  • Pinkner JS, Remaut H, Buelens F, Miller E, Aberg V, Pemberton N, Hendenström M, Larsson A, Seed P et al (2006) Rationally designed small compounds inhibit pilus biogenesis in uropathogenic bacteria. Proc Natl Acad Sci USA 103:17897–17902

    Article  CAS  PubMed  Google Scholar 

  • Saulino ET, Bullitt E, Hultgren SJ (2000) Snapshots of usher-mediated protein secretion and ordered pilus assembly. Proc Natl Acad Sci USA 97:9240–9245

    Article  CAS  PubMed  Google Scholar 

  • Svensson A, Larsson A, Emtenäs H, Hedenström M, Fex T, Hultgren SJ, Pinkner JS, Almqvist F, Kihlberg J (2001) Design and evaluation of pilicides: potential novel antibacterial agents directed against uropathogenic Escherichia coli. Chembiochem. 2:915–918

    Article  CAS  PubMed  Google Scholar 

  • Thanassi DG, Stathopoulos C, Dodson K, Geiger D, Hultgren SJ (2002) Bacterial outer membrane ushers contain distinct targeting and assembly domains for pilus biogenesis. J Bacteriol 184:6260–6269

    Article  CAS  PubMed  Google Scholar 

  • Van Loy C, Sokurenko EV, Moseley SL (2002a) The major structural subunits of Dr and F1845 fimbriae are adhesins. Infect Immun 70:1694–1702

    Article  PubMed  CAS  Google Scholar 

  • Van Loy C, Sokurenko EV, Samudrala R, Moseley SL (2002b) Identification of amino acids in the Dr adhesin required for binding to decay-accelerating factor. Mol Microbiol 45:439–452

    Article  PubMed  Google Scholar 

  • Veselovsky AV, Ivanov YP, Ivanov AS, Archakov AI, Lewi P, Janssen P (2002) Protein-protein interactions: mechanisms and modification by drugs. J Mol Recognit 15:405–422

    Article  CAS  PubMed  Google Scholar 

  • Westerlund B, Kuusela P, Risteli J, Risteli L, Vartio T, Rauvala H, Virkola R, Korhonen TK (1989) The O75X adhesin of uropathogenic Escherichia coli is a type IV collagen-binding protein. Mol Microbiol 3:329–337

    Article  CAS  PubMed  Google Scholar 

  • Zalewska B, Piątek R, Konopa G, Nowicki B, Nowicki S, Kur J (2003) Chiemric Dr fimbriae with a Herpes Simplex virus type 1 epitope as a model for a recombinant vaccine. Infect Immun 71:5505–5513

    Article  CAS  PubMed  Google Scholar 

  • Zalewska B, Piątek R, Bury K, Samet B, Nowicki BJ, Nowicki S, Kur J (2005) A surface-exposed DraD protein of uropathogenic Escherichia coli bearing Dr fimbriae may be expressed and secreted independently from DraC usher and DraE adhesin. Microbiol 151:2477–2486

    Article  CAS  Google Scholar 

  • Zavialov A, Zav’yalova G, Korpela T, Zav’yalov V (2007) FGL chaperone-assembled fimbrial polyadhesins: anti-immune armament of Gram-negative bacterial pathogens. FEMS Microbiol Rev 31:478–514

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was supported by the Polish State Committee for Scientific Research, grants N301 11631/3511 to JK.

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Authors and Affiliations

  1. Department of Microbiology, Gdańsk University of Technology, ul. G. Narutowicza 11/12, 80-952, Gdańsk, Poland

    Beata Zalewska-Piątek, Marta Kur, Sabina Wilkanowicz, Rafał Piątek & Józef Kur

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  1. Beata Zalewska-Piątek
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  2. Marta Kur
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  3. Sabina Wilkanowicz
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  4. Rafał Piątek
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  5. Józef Kur
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Correspondence to Beata Zalewska-Piątek.

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Communicated by Jorge Membrillo-Hernández.

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Zalewska-Piątek, B., Kur, M., Wilkanowicz, S. et al. The DraC usher in Dr fimbriae biogenesis of uropathogenic E. coli Dr+ strains. Arch Microbiol 192, 351–363 (2010). https://doi.org/10.1007/s00203-010-0564-x

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  • DOI: https://doi.org/10.1007/s00203-010-0564-x

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