Structure of polar pili from Pseudomonas aeruginosa strains K and O

Folkhard, W.; Marvin, D.A.; Watts, Tania H.; Paranchych, W.

doi:10.1016/0022-2836(81)90261-8

Cited by 100 publications

(70 citation statements)

References 26 publications

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“…Unfortunately, because of insolubility and purification problems, fimbrial subunits have been difficult to crystallize. Despite the fact that the three-dimensional structure of whole fimbriae is well documented (Folkhard et al, 1981;Bullitt and Makowski, 1995;Simons et al, 1994), pilin of Neisseria gonorrhoeae is the only fimbrial subunit to be crystallized and solved successfully at the atomic level (Parge et al, 1995). In the absence of X-ray crystallogaphy information, site-directed mutagenesis should continue to be an informative approach to define structure-function relationships in the Dr family of adhesins.…”

Section: Discussionmentioning

confidence: 99%

**Mutational analysis of receptor binding mediated by the Dr family of Escherichia coli adhesins**

Carnoy

Moseley

1997

Molecular Microbiology

107

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SummaryThe fimbrial and afimbrial adhesins of the Dr family mediate the adherence of uropathogenic and diarrhoea-associated Escherichia coli to decay-accelerating factor (DAF) present on erythrocytes and other cell types. The Dr haemagglutinin binds type IV collagen and, unlike other members of the Dr family, mediates an adherence inhibited in the presence of chloramphenicol. We examined the ability of other members of the Dr family-AFAI, AFAIII, and F1845-to bind to type IV collagen, and demonstrated that the collagen-binding phenotype was unique to the Dr haemagglutinin. We employed site-directed mutagenesis to demonstrate the requirement of a negatively charged amino-acid at position 54 of the Dr haemagglutinin subunit for chloramphenicol sensitivity of binding. Mutations at position 32, 40, 54, 90, and 113 differently affected type IV collagen binding and chloramphenicol sensitivity of binding, while retaining DAF-binding capability. These results suggest the existence of a conformational receptor-binding domain in the major structural subunit of Dr family adhesins and demonstrate that chloramphenicol sensitivity of binding and adherence to type IV collagen were independent and separable phenotypes. Finally, we showed that the two conserved cysteine residues of Dr family structural subunits form a disulphide bond and that mutations of these residues abolish haemagglutination and binding to type IV collagen.

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Section: Discussionmentioning

confidence: 99%

**Mutational analysis of receptor binding mediated by the Dr family of Escherichia coli adhesins**

Carnoy

Moseley

1997

Molecular Microbiology

107

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“…They are primarily composed of a small (145-160 aa) structural subunit (pilin or PilA in P. aeruginosa) with a characteristic highly conserved and highly hydrophobic aminoterminal region. This forms the core of the helical structure, whose outer face is comprised of the more hydrophilic and more variable domains of the subunit (Folkhard et al, 1981 ;Paranchych & Frost, 1988 ;Parge et al, 1990 ;Forest & Tainer, 1997).…”

Section: Introductionmentioning

confidence: 99%

Identification of a novel gene, fimV, involved in twitching motility in Pseudomonas aeruginosa The GenBank accession number for the sequence determined in this work is U93274.

et al. 2000

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Transposon mutagenesis was used to identify a new locus required for twitching motility in Pseudomonas aeruginosa. Four Tn5-B21 mutants which lacked twitching motility and a fifth which exhibited impaired motility were found to map to the same KpnI restriction fragment at approximately 40 min on the P. aeruginosa genome. Cloning and sequencing studies showed that all five transposon insertions occurred within the same 28 kb ORF, which was termed fimV. The product of this gene has a putative peptidoglycan-binding domain, predicted transmembrane domains, a highly acidic C terminus and anomalous electrophoretic migration, indicating unusual primary or secondary structure. The P. aeruginosa genome also possesses a paralogue of fimV. Homologues of fimV were also found in the sequenced genomes of the other type-IV-fimbriated bacteria Neisseria gonorrhoeae, Neisseria meningitidis, Legionella pneumophila and Vibrio cholerae, but not in those of other bacteria which lack type IV fimbriae. A fimV homologue was also found in the genome sequence of Shewanella putrefaciens, along with many other homologues of type IV fimbrial genes, indicating that this bacterium is also likely to produce type IV fimbriae. Wild-type twitching motility was restored to fimV mutants by complementation in a dosage-dependent manner. Overexpression of fimV resulted in an unusual phenotype where the cells were massively elongated and migrated in large convoys at the periphery of the colony. It is suggested that FimV may be involved in remodelling of the peptidoglycan layer to enable assembly of the type IV fimbrial structure and machinery.

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“…The arrangement of subunits in fimbrial fibers has been examined, using optical or X-ray diffraction (3,7,8,24,26). These experiments revealed that the morphological differences between the thick and thin fimbriae are in the arrangement of the subunits.…”

Section: Amino Acid Analysis Of the Us5 Fimbriaementioning

confidence: 99%

“…These experiments revealed that the morphological differences between the thick and thin fimbriae are in the arrangement of the subunits. In the thick fimbriae, the subunits are packed in a helical manner leaving a central hole in a fimbrial filament (7,8,27). On the other hand, in the thin fimbriae, the subunits are also arranged helically, but there is no central hole (24).…”

Section: Amino Acid Analysis Of the Us5 Fimbriaementioning

confidence: 99%

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Antigenic Determinants on Fimbriae of Serratia marcescens US5 Analyzed Using Monoclonal Antibodies

Jingushi

Mitsuyama

Moriya

et al. 1987

Microbiology and Immunology

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The antigenic sites on small thin fimbriae of Serratia marcescens strain US5 were investigated using immunoelectron microscopy and monoclonal antibodies (MAbs). Negative staining of the fimbriae after treatment with MAbs showed a regularly spaced arrangement of the antibody molecules. When the subunit peptide was subjected to immunoblotting using the MAbs, a single band with a molecular weight of approximately 19kD was evident. This binding of the MAbs to the subunit peptide was completely abrogated after treatment with 2-mercaptoethanol, thereby suggesting the important role of disulfide linkage in the maintenance of the conformation of the antigenic site reacted with MAbs. Amino acid analysis of the subunit peptide revealed two cysteine residues, and cysteine residues were absent in the N-terminal portion.Bacterial fimbriae or pili are important virulent factors mediating adhesion of bacteria to the mucosal surface. Inhibition of the adherence by antibodies against fimbriae is an effective approach to prevent bacterial infection, in the early stage (17) .Chemical analyses of fimbriae showed that several different functional regions were involved (5,18,20,24). In the fimbriae of Neisseria gonorrheae, the region for adherence is located close to the antigenic site, and type-specific antibodies can effectively inhibit the adherence (24, 27). Morphologically, fimbriae can be grouped into two types, large thick fimbriae and small thin ones (13). Each fimbrial fiber consists of subunit peptides with a molecular weight of approximately 20,000 daltons. X-ray and optical diffraction studies revealed that the fimbrial fiber is formed by helical arrangement of subunits (7,8,27). Differences in the size of the fimbriae are mainly dependent on the mode of packing of the subunits, in a helical fashion (13). The relation of the structure and the functional regions of the fimbrial subunits has not been elucidated.Serratia marcescens causes nosocomial infections. We reported that S. marcescens strain US5 possessed mannose-sensitive fimbriae, that the adherence of this bacteria to the urinary bladder mucosal surface was mediated by small thin fimbriae, and that anti-fimbriae sera effectively inhibited the adherence (15,28). In the present 879

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Structure of polar pili from Pseudomonas aeruginosa strains K and O

Cited by 100 publications

References 26 publications

**Mutational analysis of receptor binding mediated by the Dr family of Escherichia coli adhesins**

**Mutational analysis of receptor binding mediated by the Dr family of Escherichia coli adhesins**

Identification of a novel gene, fimV, involved in twitching motility in Pseudomonas aeruginosa The GenBank accession number for the sequence determined in this work is U93274.

Antigenic Determinants on Fimbriae of Serratia marcescens US5 Analyzed Using Monoclonal Antibodies

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