Mutation of the conserved calcium-binding motif in Neisseria gonorrhoeae PilC1 impacts adhesion but not piliation

Yuan Cheng, Michael D.L. Johnson, Christine Burillo-Kirch, Jeffrey C. Mocny, James E. Anderson, Christopher K. Garrett, Matthew R. Redinbo, Christopher E. Thomas

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

Neisseria gonorrhoeae PilC1 is a member of the PilC family of type IV pilus-associated adhesins found in Neisseria species and other type IV pilus-producing genera. Previously, a calcium-binding domain was described in the C-terminal domains of PilY1 of Pseudomonas aeruginosa and in PilC1 and PilC2 of Kingella kingae. Genetic analysis of N. gonorrhoeae revealed a similar calcium-binding motif in PilC1. To evaluate the potential significance of this calcium-binding region in N. gonorrhoeae, we produced recombinant full-length PilC1 and a PilC1 C-terminal domain fragment. We show that, while alterations of the calciumbinding motif disrupted the ability of PilC1 to bind calcium, they did not grossly affect the secondary structure of the protein. Furthermore, we demonstrate that both full-length wild-type PilC1 and full-length calcium-binding-deficient PilC1 inhibited gonococcal adherence to cultured human cervical epithelial cells, unlike the truncated PilC1 C-terminal domain. Similar to PilC1 in K. kingae, but in contrast to the calcium-binding mutant of P. aeruginosa PilY1, an equivalent mutation in N. gonorrhoeae PilC1 produced normal amounts of pili. However, the N. gonorrhoeae PilC1 calcium-binding mutant still had partial defects in gonococcal adhesion to ME180 cells and genetic transformation, which are both essential virulence factors in this human pathogen. Thus, we conclude that calcium binding to PilC1 plays a critical role in pilus function in N. gonorrhoeae.

Original languageEnglish (US)
Pages (from-to)4279-4289
Number of pages11
JournalInfection and Immunity
Volume81
Issue number11
DOIs
StatePublished - Nov 2013

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Immunology
  • Infectious Diseases

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    Cheng, Y., Johnson, M. D. L., Burillo-Kirch, C., Mocny, J. C., Anderson, J. E., Garrett, C. K., Redinbo, M. R., & Thomas, C. E. (2013). Mutation of the conserved calcium-binding motif in Neisseria gonorrhoeae PilC1 impacts adhesion but not piliation. Infection and Immunity, 81(11), 4279-4289. https://doi.org/10.1128/IAI.00493-13