Suppression of an absolute defect in Type IV pilus biogenesis by loss-of-function mutations in
            <i>pilT</i>
            , a twitching motility gene in
            <i>Neisseria gonorrhoeae</i>

Wolfgang, Matthew C.; Park, Hae Sun; Hayes, Stanley F.; Putten, J. P. M. Van; Koomey, Michael

doi:10.1073/pnas.95.25.14973

Cited by 130 publications

(162 citation statements)

References 29 publications

Supporting

Mentioning

150

Contrasting

Order By: Relevance

“…We found that four pili variants containing different PilE sequences (38) bound C4BP with similar apparent affinities (not shown). PilC is a 110-kDa pilus biogenesis protein that seems to be involved in the interaction between pili and MCP, because it was possible to select or genetically construct adherence negative mutants that expressed pili without detectable amounts of PilC (39,40). The results of an overlay assay suggested that C4BP interacts with the PilC subunit of pili.…”

Section: Discussionmentioning

confidence: 99%

A Novel Interaction Between Type IV Pili ofNeisseria gonorrhoeaeand the Human Complement Regulator C4b-Binding Protein

Blom

Rytkönen

Vásquez

et al. 2001

The Journal of Immunology

View full text Add to dashboard Cite

C4b-binding protein (C4BP) is an important plasma inhibitor of the classical pathway of complement activation. Several bacterial pathogens bind C4BP, which may contribute to their virulence. In the present report we demonstrate that isolated type IV pili from Neisseria gonorrhoeae bind human C4BP in a dose-dependent and saturable manner. C4BP consists of seven identical α-chains and one β-chain linked together with disulfide bridges. We found that pili bind to the α-chain of C4BP, which is composed of eight homologous complement control protein (CCP) domains. From the results of an inhibition assay with C4b and a competition assay in which we tested mutants of C4BP lacking individual CCPs, we concluded that the binding area for pili is localized to CCP1 and CCP2 of the α-chain. The binding between pili and C4BP was abolished at 0.25 M NaCl, implying that it is based mostly on ionic interactions, similarly to what have been observed for C4b-C4BP binding. Furthermore, the N-terminal part of PilC, a structural component of pili, appeared to be responsible for binding of C4BP. Membrane cofactor protein, previously shown to be a receptor for pathogenic N. gonorrhoeae on the surface of epithelial cells, competed with C4BP for binding to pili only at high concentrations, suggesting that different parts of pili are involved in these two interactions. Accordingly, high concentrations of C4BP were required to inhibit binding of N. gonorrhoeae to Chang conjunctiva cells, and no inhibition of binding was observed with cervical epithelial cells.

show abstract

Section: Discussionmentioning

confidence: 99%

A Novel Interaction Between Type IV Pili ofNeisseria gonorrhoeaeand the Human Complement Regulator C4b-Binding Protein

Blom

Rytkönen

Vásquez

et al. 2001

The Journal of Immunology

View full text Add to dashboard Cite

show abstract

“…Four of these clusters are found on Chr I of all of the sequenced Vibrio species. They include a mannose-sensitive hemagglutinin (MshA), a homolog of the PilT pilus involved in twitching motility (22), and two Vibrio PilA homologs (21,23). Two of the remaining four pilus gene clusters are paralogs (one on each V. fischeri chromosome) that are homologous to the Flp1 type IV-B tight adhesion (tad) pilus family.…”

Section: Resultsmentioning

confidence: 99%

Complete genome sequence of Vibrio fischeri : A symbiotic bacterium with pathogenic congeners

Ruby

Urbanowski

Campbell

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

330

238

View full text Add to dashboard Cite

Vibrio fischeri belongs to the Vibrionaceae, a large family of marine ␥-proteobacteria that includes several dozen species known to engage in a diversity of beneficial or pathogenic interactions with animal tissue. Among the small number of pathogenic Vibrio species that cause human diseases are Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus, the only members of the Vibrionaceae that have had their genome sequences reported. Nonpathogenic members of the genus Vibrio, including a number of beneficial symbionts, make up the majority of the Vibrionaceae, but none of these species has been similarly examined. Here we report the genome sequence of V. fischeri ES114, which enters into a mutualistic symbiosis in the light organ of the bobtail squid, Euprymna scolopes. Analysis of this sequence has revealed surprising parallels with V. cholerae and other pathogens.genomics ͉ pili ͉ symbiosis ͉ toxins ͉ toxin-coregulated pilus

show abstract

“…Procedures for SDS͞ PAGE and immunoblotting have been described previously (13). Quantitative pilus purification was carried out as previously described (13).…”

Section: Characterization Of Twitching Motility and Competence For Namentioning

confidence: 99%

“…The conclusion that PilC is a neisserial Tfp-associated adhesin is based primarily on the findings that (i) purified recombinant PilC imparts human epithelial cell binding activity on latex beads to which it has been coupled and blocks binding of piliated organisms to human epithelial cells (7) and (ii) piliated mutants lacking PilC fail to adhere (12,13). However, several studies have suggested a role for the pilin subunit PilE in receptor recognition.…”

mentioning

confidence: 99%

Neisseria gonorrhoeae PilV, a type IV pilus-associated protein essential to human epithelial cell adherence

Winther‐Larsen

Hegge

Wolfgang

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

121

136

View full text Add to dashboard Cite

Type IV pili (Tfp) of Neisseria gonorrhoeae, the Gram-negative etiologic agent of gonorrhea, facilitate colonization of the human host. Tfp are assumed to play a key role in the initial adherence to human epithelial cells by virtue of the associated adhesin protein PilC. To examine the structural and functional basis for adherence in more detail, we identified potential genes encoding polypeptides sharing structural similarities to PilE (the Tfp subunit) within the N. gonorrhoeae genome sequence database. We show here that a fiber subunit-like protein, termed PilV, is essential to organelle-associated adherence but dispensable for Tfp biogenesis and other pilus-related phenotypes, including autoagglutination, competence for natural transformation, and twitching motility. The adherence defect in pilV mutants cannot be attributed to reduced levels of piliation, defects in fiber anchoring to the bacterial cell surface, or to unstable pilus expression related to organelle retraction. PilV is expressed at low levels relative to PilE and copurifies with Tfp fibers in a PilC-dependent fashion. Purified Tfp from pilV mutants contain PilC adhesin at reduced levels. Taken together, these data support a model in which PilV functions in adherence by promoting the functional display of PilC in the context of the pilus fiber.

show abstract

Suppression of an absolute defect in Type IV pilus biogenesis by loss-of-function mutations in pilT , a twitching motility gene in Neisseria gonorrhoeae

Cited by 130 publications

References 29 publications

A Novel Interaction Between Type IV Pili ofNeisseria gonorrhoeaeand the Human Complement Regulator C4b-Binding Protein

A Novel Interaction Between Type IV Pili ofNeisseria gonorrhoeaeand the Human Complement Regulator C4b-Binding Protein

Complete genome sequence of Vibrio fischeri : A symbiotic bacterium with pathogenic congeners

Neisseria gonorrhoeae PilV, a type IV pilus-associated protein essential to human epithelial cell adherence

Contact Info

Product

Resources

About