Recognition of the N‐terminal lectin domain of FimH adhesin by the usher FimD is required for type 1 pilus biogenesis

In this work we discover that a specific recognition of the N‐terminal lectin domain of FimH adhesin by the usher FimD is essential for the biogenesis of type 1 pili in Escherichia coli. These filamentous organelles are assembled by the chaperone–usher pathway, in which binary complexes between fimbrial subunits and the periplasmic chaperone FimC are recognized by the outer membrane protein FimD (the usher). FimH adhesin initiates fimbriae polymerization and is the first subunit incorporated in the filament. Accordingly, FimD shows higher affinity for the FimC/FimH complex although the structural basis of this specificity is unknown. We have analysed the assembly into fimbria, and the interaction with FimD in vivo, of FimH variants in which the N‐terminal lectin domain of FimH was deleted or substituted by different immunoglobulin (Ig) domains, or in which these Ig domains were fused to the N‐terminus of full‐length FimH. From these data, along with the analysis of a FimH mutant with a single amino acid change (G16D) in the N‐terminal lectin domain, we conclude that the lectin domain of FimH is recognized by FimD usher as an essential step for type 1 pilus biogenesis.

[1]  L. Wyns,et al.  The affinity of the FimH fimbrial adhesin is receptor-driven and quasi-independent of Escherichia coli pathotypes , 2006, Molecular microbiology.

[2]  R. Glockshuber,et al.  Mechanism of fibre assembly through the chaperone–usher pathway , 2006, EMBO reports.

[3]  J. Kormanec,et al.  Pushing the envelope: extracytoplasmic stress responses in bacterial pathogens , 2006, Nature Reviews Microbiology.

[4]  D. Thanassi,et al.  Analysis of the requirements for pilus biogenesis at the outer membrane usher and the function of the usher C‐terminus , 2006, Molecular microbiology.

[5]  T. Ratliff.  Receptor binding studies disclose a novel class of high-affinity inhibitors of the Escherichia coli FimH adhesin. , 2005, The Journal of urology.

[6]  P. Hudson,et al.  Engineered antibody fragments and the rise of single domains , 2005, Nature Biotechnology.

[7]  R. Glockshuber,et al.  Structural basis of chaperone–subunit complex recognition by the type 1 pilus assembly platform FimD , 2005, The EMBO journal.

[8]  T. Silhavy,et al.  Sensing external stress: watchdogs of the Escherichia coli cell envelope. , 2005, Current opinion in microbiology.

[9]  N. Henderson,et al.  Topology of the Outer Membrane Usher PapC Determined by Site-directed Fluorescence Labeling* , 2004, Journal of Biological Chemistry.

[10]  Huilin Li,et al.  The outer membrane usher forms a twin-pore secretion complex. , 2004, Journal of molecular biology.

[11]  G. Waksman,et al.  Fiber assembly by the chaperone-usher pathway. , 2004, Biochimica et biophysica acta.

[12]  D. Thanassi,et al.  The Usher N Terminus Is the Initial Targeting Site for Chaperone-Subunit Complexes and Participates in Subsequent Pilus Biogenesis Events , 2004, Journal of bacteriology.

[13]  V. de Lorenzo,et al.  Secretion of proteins with dimerization capacity by the haemolysin type I transport system of Escherichia coli , 2004, Molecular microbiology.

[14]  V. de Lorenzo,et al.  Structural tolerance of bacterial autotransporters for folded passenger protein domains , 2004, Molecular microbiology.

[15]  Lode Wyns,et al.  The fimbrial adhesin F17‐G of enterotoxigenic Escherichia coli has an immunoglobulin‐like lectin domain that binds N‐acetylglucosamine , 2003, Molecular microbiology.

[16]  R. Glockshuber,et al.  Identification and characterization of the chaperone-subunit complex-binding domain from the type 1 pilus assembly platform FimD. , 2003, Journal of molecular biology.

[17]  J. Pinkner,et al.  Chaperone-Subunit-Usher Interactions Required for Donor Strand Exchange during Bacterial Pilus Assembly , 2003, Journal of bacteriology.

[18]  Christos Stathopoulos,et al.  Bacterial Outer Membrane Ushers Contain Distinct Targeting and Assembly Domains for Pilus Biogenesis , 2002, Journal of bacteriology.

[19]  G. Waksman,et al.  Chaperone Priming of Pilus Subunits Facilitates a Topological Transition that Drives Fiber Formation , 2002, Cell.

[20]  Ueli Aebi,et al.  Exploring the 3D molecular architecture of Escherichia coli type 1 pili. , 2002, Journal of molecular biology.

[21]  R. Glockshuber,et al.  Chaperone-independent folding of type 1 pilus domains. , 2002, Journal of molecular biology.

[22]  V. de Lorenzo,et al.  Production of functional single-chain Fv antibodies in the cytoplasm of Escherichia coli. , 2002, Journal of molecular biology.

[23]  M. Mulvey,et al.  Adhesion and entry of uropathogenic Escherichia coli , 2002, Cellular microbiology.

[24]  G. Waksman,et al.  Structural basis of tropism of Escherichia coli to the bladder during urinary tract infection , 2002, Molecular microbiology.

[25]  L. Wyns,et al.  Single‐domain antibody fragments with high conformational stability , 2002, Protein science : a publication of the Protein Society.

[26]  S. Hultgren,et al.  Requirement of Rho‐family GTPases in the invasion of Type 1‐piliated uropathogenic Escherichia coli , 2002, Cellular microbiology.

[27]  R. Glockshuber,et al.  Uroplakin Ia is the urothelial receptor for uropathogenic Escherichia coli: evidence from in vitro FimH binding. , 2001, Journal of cell science.

[28]  S E Hufton,et al.  Building novel binding ligands to B7.1 and B7.2 based on human antibody single variable light chain domains. , 2001, Journal of molecular biology.

[29]  S. Muyldermans,et al.  Single domain camel antibodies: current status. , 2001, Journal of biotechnology.

[30]  L. Wyns,et al.  Recognition of antigens by single-domain antibody fragments: the superfluous luxury of paired domains. , 2001, Trends in biochemical sciences.

[31]  A. Plückthun,et al.  Stability engineering of antibody single-chain Fv fragments. , 2001, Journal of molecular biology.

[32]  M. Schembri,et al.  Fimbrial surface display systems in bacteria: from vaccines to random libraries. , 2000, Microbiology.

[33]  S. Hultgren,et al.  Snapshots of usher-mediated protein secretion and ordered pilus assembly. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[34]  S. Hultgren,et al.  Multiple pathways allow protein secretion across the bacterial outer membrane. , 2000, Current opinion in cell biology.

[35]  H. Hasman,et al.  Expression and purification of the mannose recognition domain of the FimH adhesin. , 2000, FEMS microbiology letters.

[36]  P. van Gelder,et al.  Structure and function of bacterial outer membrane proteins: barrels in a nutshell , 2000, Molecular microbiology.

[37]  J. Pinkner,et al.  Type 1 pilus‐mediated bacterial invasion of bladder epithelial cells , 2000, The EMBO journal.

[38]  V. de Lorenzo,et al.  Probing secretion and translocation of a β‐autotransporter using a reporter single‐chain Fv as a cognate passenger domain , 1999, Molecular microbiology.

[39]  V. Stojanoff,et al.  X-ray structure of the FimC-FimH chaperone-adhesin complex from uropathogenic Escherichia coli. , 1999, Science.

[40]  A. Plückthun,et al.  Different equilibrium stability behavior of ScFv fragments: identification, classification, and improvement by protein engineering. , 1999, Biochemistry.

[41]  T. Silhavy,et al.  The sigmaE and Cpx regulatory pathways: overlapping but distinct envelope stress responses. , 1999, Current opinion in microbiology.

[42]  S. Hultgren,et al.  Induction and evasion of host defenses by type 1-piliated uropathogenic Escherichia coli. , 1998, Science.

[43]  M. Schembri,et al.  Heterobinary Adhesins Based on theEscherichia coli FimH Fimbrial Protein , 1998, Applied and Environmental Microbiology.

[44]  J. Pinkner,et al.  Ramifications of kinetic partitioning on usher‐mediated pilus biogenesis , 1998, The EMBO journal.

[45]  S. Hultgren,et al.  The chaperone/usher pathway: a major terminal branch of the general secretory pathway. , 1998, Current opinion in microbiology.

[46]  S. Hultgren,et al.  The PapC usher forms an oligomeric channel: implications for pilus biogenesis across the outer membrane. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[47]  S. Langermann,et al.  Prevention of mucosal Escherichia coli infection by FimH-adhesin-based systemic vaccination. , 1997, Science.

[48]  C. Svanborg,et al.  Type 1 fimbrial expression enhances Escherichia coli virulence for the urinary tract. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[49]  M. Schembri,et al.  Linker insertion analysis of the FimH adhesin of type 1 fimbriae in an Escherichia coli fimH-null background. , 1996, FEMS microbiology letters.

[50]  L. Poulsen,et al.  Chimeric FimH adhesin of type 1 fimbriae: a bacterial surface display system for heterologous sequences. , 1995, Microbiology.

[51]  J. Pinkner,et al.  FimH adhesin of type 1 pili is assembled into a fibrillar tip structure in the Enterobacteriaceae. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[52]  P. Klemm,et al.  The export systems of type 1 and F1C fimbriae are interchangeable but work in parental pairs , 1995, Journal of bacteriology.

[53]  P Bork,et al.  The immunoglobulin fold. Structural classification, sequence patterns and common core. , 1994, Journal of molecular biology.

[54]  R. Marre,et al.  Reciprocal exchange of minor components of type 1 and F1C fimbriae results in hybrid organelles with changed receptor specificities , 1994, Journal of bacteriology.

[55]  F. Jacob-Dubuisson,et al.  Outer-membrane PapC molecular usher discriminately recognizes periplasmic chaperone-pilus subunit complexes. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[56]  S. Garges.  A Short Course in Bacterial Genetics. A Laboratory Manual and Handbook for Escherichia coli and Related Bacteria. By Jeffrey H. Miller. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 1992. , 1993 .

[57]  V. de Lorenzo,et al.  Analysis of Pseudomonas gene products using lacIq/Ptrp-lac plasmids and transposons that confer conditional phenotypes. , 1993, Gene.

[58]  C. Brändén,et al.  Interactive surface in the PapD chaperone cleft is conserved in pilus chaperone superfamily and essential in subunit recognition and assembly. , 1992, The EMBO journal.

[59]  C. Bloch,et al.  A key role for type 1 pili in enterobacterial communicability , 1992, Molecular microbiology.

[60]  M. S. McClain,et al.  Type 1 fimbriae mutants of Escherichia coli K12: characterization of recognized afimbriate strains and construction of new fim deletion mutants , 1991, Molecular microbiology.

[61]  K. Krogfelt,et al.  Direct evidence that the FimH protein is the mannose-specific adhesin of Escherichia coli type 1 fimbriae , 1990, Infection and immunity.

[62]  C. Bloch,et al.  Impaired colonization by and full invasiveness of Escherichia coli K1 bearing a site-directed mutation in the type 1 pilin gene , 1990, Infection and immunity.

[63]  K. Krogfelt,et al.  Investigation of minor components of Escherichia coli type 1 fimbriae: protein chemical and immunological aspects. , 1988, Microbial pathogenesis.

[64]  J. Dunn,et al.  ompT encodes the Escherichia coli outer membrane protease that cleaves T7 RNA polymerase during purification , 1988, Journal of bacteriology.

[65]  P. Klemm,et al.  Three fim genes required for the regulation of length and mediation of adhesion of Escherichia coli type 1 fimbriae , 1987, Molecular and General Genetics MGG.

[66]  A. Schaeffer,et al.  Regulation of production of type 1 pili among urinary tract isolates of Escherichia coli , 1986, Infection and Immunity.

[67]  A. Schaeffer,et al.  Role of type 1 pili and effects of phase variation on lower urinary tract infections produced by Escherichia coli , 1985, Infection and immunity.

[68]  N. Sharon,et al.  Adherence of Escherichia coli to human mucosal cells mediated by mannose receptors , 1977, Nature.

[69]  D. Old.  Inhibition of the interaction between fimbrial haemagglutinins and erythrocytes by D-mannose and other carbohydrates. , 1972, Journal of general microbiology.

[70]  H. Boyer,et al.  A complementation analysis of the restriction and modification of DNA in Escherichia coli. , 1969, Journal of molecular biology.

[71]  P. Klemm,et al.  The fimD gene required for cell surface localization of Escherichia coli type 1 fimbriae , 2004, Molecular and General Genetics MGG.

[72]  D. Thanassi.  Ushers and secretins: channels for the secretion of folded proteins across the bacterial outer membrane. , 2002, Journal of molecular microbiology and biotechnology.

[73]  S. Hultgren,et al.  Assembly of complex organelles: pilus biogenesis in gram-negative bacteria as a model system. , 2000, Methods.

[74]  M. Schembri,et al.  Bioaccumulation of heavy metals by fimbrial designer adhesins. , 1999, FEMS microbiology letters.

[75]  V. de Lorenzo,et al.  Analysis and construction of stable phenotypes in gram-negative bacteria with Tn5- and Tn10-derived minitransposons. , 1994, Methods in enzymology.

[76]  J. Steitz,et al.  Identification of a sex-factor-affinity site in E. coli as gamma delta. , 1981, Cold Spring Harbor symposia on quantitative biology.