Abstract
The Gram-negative bacterial pathogen Neisseria gonorrhoeae is naturally competent for transformation with species-related DNA. We show here that two phase-variable pilus-associated proteins, the major pilus subunit (pilin, or PilE) and PilC, a factor known to function in the assembly and adherence of gonococcal pili, are essential for transformation competence. The PilE and PilC proteins are necessary for the conversion of linearized plasmid DNA carrying the Neisseria-specific DNA uptake signal into a DNase-resistant form. The biogenesis of typical pilus fibers is neither essential nor sufficient for this process. DNA uptake deficiency of defined piliated pilC1,2 double mutants can be complemented by expression of a cloned pilC2 gene in trans. The PilC defect can also be restored by the addition of purified PilC protein, or better, pili containing PilC protein, to the mutant gonococci. Our data suggest that the two phase-variable Pil proteins act on the bacterial cell surface and cooperate in DNA recognition and/or outer membrane translocation.
Full text
PDF




Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Biswas G. D., Burnstein K. L., Sparling P. F. Linearization of donor DNA during plasmid transformation in Neisseria gonorrhoeae. J Bacteriol. 1986 Nov;168(2):756–761. doi: 10.1128/jb.168.2.756-761.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Biswas G. D., Lacks S. A., Sparling P. F. Transformation-deficient mutants of piliated Neisseria gonorrhoeae. J Bacteriol. 1989 Feb;171(2):657–664. doi: 10.1128/jb.171.2.657-664.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Biswas G. D., Sox T., Blackman E., Sparling P. F. Factors affecting genetic transformation of Neisseria gonorrhoeae. J Bacteriol. 1977 Feb;129(2):983–992. doi: 10.1128/jb.129.2.983-992.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dubnau D. Genetic competence in Bacillus subtilis. Microbiol Rev. 1991 Sep;55(3):395–424. doi: 10.1128/mr.55.3.395-424.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Elkins C., Thomas C. E., Seifert H. S., Sparling P. F. Species-specific uptake of DNA by gonococci is mediated by a 10-base-pair sequence. J Bacteriol. 1991 Jun;173(12):3911–3913. doi: 10.1128/jb.173.12.3911-3913.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Facius D., Meyer T. F. A novel determinant (comA) essential for natural transformation competence in Neisseria gonorrhoeae and the effect of a comA defect on pilin variation. Mol Microbiol. 1993 Nov;10(4):699–712. doi: 10.1111/j.1365-2958.1993.tb00942.x. [DOI] [PubMed] [Google Scholar]
- Feavers I. M., Heath A. B., Bygraves J. A., Maiden M. C. Role of horizontal genetic exchange in the antigenic variation of the class 1 outer membrane protein of Neisseria meningitidis. Mol Microbiol. 1992 Feb;6(4):489–495. doi: 10.1111/j.1365-2958.1992.tb01493.x. [DOI] [PubMed] [Google Scholar]
- Frosch M., Meyer T. F. Transformation-mediated exchange of virulence determinants by co-cultivation of pathogenic Neisseriae. FEMS Microbiol Lett. 1992 Dec 15;100(1-3):345–349. doi: 10.1111/j.1574-6968.1992.tb14062.x. [DOI] [PubMed] [Google Scholar]
- Gibbs C. P., Reimann B. Y., Schultz E., Kaufmann A., Haas R., Meyer T. F. Reassortment of pilin genes in Neisseria gonorrhoeae occurs by two distinct mechanisms. Nature. 1989 Apr 20;338(6217):651–652. doi: 10.1038/338651a0. [DOI] [PubMed] [Google Scholar]
- Goodman S. D., Scocca J. J. Identification and arrangement of the DNA sequence recognized in specific transformation of Neisseria gonorrhoeae. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6982–6986. doi: 10.1073/pnas.85.18.6982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Haas R., Schwarz H., Meyer T. F. Release of soluble pilin antigen coupled with gene conversion in Neisseria gonorrhoeae. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9079–9083. doi: 10.1073/pnas.84.24.9079. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Halter R., Pohlner J., Meyer T. F. Mosaic-like organization of IgA protease genes in Neisseria gonorrhoeae generated by horizontal genetic exchange in vivo. EMBO J. 1989 Sep;8(9):2737–2744. doi: 10.1002/j.1460-2075.1989.tb08415.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hobbs M., Mattick J. S. Common components in the assembly of type 4 fimbriae, DNA transfer systems, filamentous phage and protein-secretion apparatus: a general system for the formation of surface-associated protein complexes. Mol Microbiol. 1993 Oct;10(2):233–243. doi: 10.1111/j.1365-2958.1993.tb01949.x. [DOI] [PubMed] [Google Scholar]
- Jonsson A. B., Nyberg G., Normark S. Phase variation of gonococcal pili by frameshift mutation in pilC, a novel gene for pilus assembly. EMBO J. 1991 Feb;10(2):477–488. doi: 10.1002/j.1460-2075.1991.tb07970.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jonsson A. B., Pfeifer J., Normark S. Neisseria gonorrhoeae PilC expression provides a selective mechanism for structural diversity of pili. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3204–3208. doi: 10.1073/pnas.89.8.3204. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kupsch E. M., Knepper B., Kuroki T., Heuer I., Meyer T. F. Variable opacity (Opa) outer membrane proteins account for the cell tropisms displayed by Neisseria gonorrhoeae for human leukocytes and epithelial cells. EMBO J. 1993 Feb;12(2):641–650. doi: 10.1002/j.1460-2075.1993.tb05697.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meyer T. F., Billyard E., Haas R., Storzbach S., So M. Pilus genes of Neisseria gonorrheae: chromosomal organization and DNA sequence. Proc Natl Acad Sci U S A. 1984 Oct;81(19):6110–6114. doi: 10.1073/pnas.81.19.6110. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meyer T. F., Pohlner J., van Putten J. P. Biology of the pathogenic Neisseriae. Curr Top Microbiol Immunol. 1994;192:283–317. doi: 10.1007/978-3-642-78624-2_13. [DOI] [PubMed] [Google Scholar]
- Pugsley A. P., d'Enfert C., Reyss I., Kornacker M. G. Genetics of extracellular protein secretion by gram-negative bacteria. Annu Rev Genet. 1990;24:67–90. doi: 10.1146/annurev.ge.24.120190.000435. [DOI] [PubMed] [Google Scholar]
- Rudel T., Scheurerpflug I., Meyer T. F. Neisseria PilC protein identified as type-4 pilus tip-located adhesin. Nature. 1995 Jan 26;373(6512):357–359. doi: 10.1038/373357a0. [DOI] [PubMed] [Google Scholar]
- Rudel T., van Putten J. P., Gibbs C. P., Haas R., Meyer T. F. Interaction of two variable proteins (PilE and PilC) required for pilus-mediated adherence of Neisseria gonorrhoeae to human epithelial cells. Mol Microbiol. 1992 Nov;6(22):3439–3450. doi: 10.1111/j.1365-2958.1992.tb02211.x. [DOI] [PubMed] [Google Scholar]
- Scocca J. J. The role of transformation in the variability of the Neisseria gonorrhoeae cell surface. Mol Microbiol. 1990 Mar;4(3):321–327. doi: 10.1111/j.1365-2958.1990.tb00599.x. [DOI] [PubMed] [Google Scholar]
- Seifert H. S., Ajioka R. S., Marchal C., Sparling P. F., So M. DNA transformation leads to pilin antigenic variation in Neisseria gonorrhoeae. Nature. 1988 Nov 24;336(6197):392–395. doi: 10.1038/336392a0. [DOI] [PubMed] [Google Scholar]
- Seifert H. S., Ajioka R. S., Paruchuri D., Heffron F., So M. Shuttle mutagenesis of Neisseria gonorrhoeae: pilin null mutations lower DNA transformation competence. J Bacteriol. 1990 Jan;172(1):40–46. doi: 10.1128/jb.172.1.40-46.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith J. M., Dowson C. G., Spratt B. G. Localized sex in bacteria. Nature. 1991 Jan 3;349(6304):29–31. doi: 10.1038/349029a0. [DOI] [PubMed] [Google Scholar]
- Smith J. M., Smith N. H., O'Rourke M., Spratt B. G. How clonal are bacteria? Proc Natl Acad Sci U S A. 1993 May 15;90(10):4384–4388. doi: 10.1073/pnas.90.10.4384. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sparling P. F. Genetic transformation of Neisseria gonorrhoeae to streptomycin resistance. J Bacteriol. 1966 Nov;92(5):1364–1371. doi: 10.1128/jb.92.5.1364-1371.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tommassen J., Filloux A., Bally M., Murgier M., Lazdunski A. Protein secretion in Pseudomonas aeruginosa. FEMS Microbiol Rev. 1992 Sep;9(1):73–90. doi: 10.1016/0378-1097(92)90336-m. [DOI] [PubMed] [Google Scholar]
- Zhang Q. Y., DeRyckere D., Lauer P., Koomey M. Gene conversion in Neisseria gonorrhoeae: evidence for its role in pilus antigenic variation. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5366–5370. doi: 10.1073/pnas.89.12.5366. [DOI] [PMC free article] [PubMed] [Google Scholar]