Skip to main content
Genetics logoLink to Genetics
. 2000 Feb;154(2):523–532. doi: 10.1093/genetics/154.2.523

A homologue of the recombination-dependent growth gene, rdgC, is involved in gonococcal pilin antigenic variation.

I J Mehr 1, C D Long 1, C D Serkin 1, H S Seifert 1
PMCID: PMC1460959  PMID: 10655208

Abstract

Neisseria gonorrhoeae pilin undergoes high-frequency changes in primary amino acid sequence that aid in the avoidance of the host immune response and alter pilus expression. The pilin amino acid changes reflect nucleotide changes in the expressed gene, pilE, which result from nonreciprocal recombination reactions with numerous silent loci, pilS. A series of mini-transposon insertions affecting pilin antigenic variation were localized to three genes in one region of the Gc chromosome. Mutational analysis with complementation showed that a Gc gene with sequence similarity to the Escherichia coli rdgC gene is involved in pilus-dependent colony phase variation and in pilin antigenic variation. Furthermore, we show that the Gc rdgC homologue is transcriptionally linked in an operon with a gene encoding a predicted GTPase. The inability to disrupt expression of this gene suggests it is an essential gene (engA, essential neisserial GTPase). While some of the transposon mutations in rdgC and insertions in the 5'-untranslated portion of engA showed a growth defect, all transposon insertions investigated conferred an aberrant cellular morphology. Complementation analysis showed that the growth deficiencies are due to the interruption of RdgC expression and not that of EngA. The requirement of RdgC for efficient pilin variation suggests a role for this protein in specialized DNA recombination reactions.

Full Text

The Full Text of this article is available as a PDF (444.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alam K. Y., Clark D. P. Molecular cloning and sequence of the thdF gene, which is involved in thiophene and furan oxidation by Escherichia coli. J Bacteriol. 1991 Oct;173(19):6018–6024. doi: 10.1128/jb.173.19.6018-6024.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Altschul S. F., Madden T. L., Schäffer A. A., Zhang J., Zhang Z., Miller W., Lipman D. J. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 1997 Sep 1;25(17):3389–3402. doi: 10.1093/nar/25.17.3389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boslego J. W., Tramont E. C., Chung R. C., McChesney D. G., Ciak J., Sadoff J. C., Piziak M. V., Brown J. D., Brinton C. C., Jr, Wood S. W. Efficacy trial of a parenteral gonococcal pilus vaccine in men. Vaccine. 1991 Mar;9(3):154–162. doi: 10.1016/0264-410x(91)90147-x. [DOI] [PubMed] [Google Scholar]
  4. Bourne H. R., Sanders D. A., McCormick F. The GTPase superfamily: conserved structure and molecular mechanism. Nature. 1991 Jan 10;349(6305):117–127. doi: 10.1038/349117a0. [DOI] [PubMed] [Google Scholar]
  5. Chaussee M. S., Wilson J., Hill S. A. Characterization of the recD gene of Neisseria gonorrhoeae MS11 and the effect of recD inactivation on pilin variation and DNA transformation. Microbiology. 1999 Feb;145(Pt 2):389–400. doi: 10.1099/13500872-145-2-389. [DOI] [PubMed] [Google Scholar]
  6. Chen X., Court D. L., Ji X. Crystal structure of ERA: a GTPase-dependent cell cycle regulator containing an RNA binding motif. Proc Natl Acad Sci U S A. 1999 Jul 20;96(15):8396–8401. doi: 10.1073/pnas.96.15.8396. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cox M. M. Relating biochemistry to biology: how the recombinational repair function of RecA protein is manifested in its molecular properties. Bioessays. 1993 Sep;15(9):617–623. doi: 10.1002/bies.950150908. [DOI] [PubMed] [Google Scholar]
  8. Fussenegger M., Facius D., Meier J., Meyer T. F. A novel peptidoglycan-linked lipoprotein (ComL) that functions in natural transformation competence of Neisseria gonorrhoeae. Mol Microbiol. 1996 Mar;19(5):1095–1105. doi: 10.1046/j.1365-2958.1996.457984.x. [DOI] [PubMed] [Google Scholar]
  9. Fussenegger M., Kahrs A. F., Facius D., Meyer T. F. Tetrapac (tpc), a novel genotype of Neisseria gonorrhoeae affecting epithelial cell invasion, natural transformation competence and cell separation. Mol Microbiol. 1996 Mar;19(6):1357–1372. doi: 10.1111/j.1365-2958.1996.tb02479.x. [DOI] [PubMed] [Google Scholar]
  10. Fyfe J. A., Davies J. K. Nucleotide sequence and expression in Escherichia coli of the recA gene of Neisseria gonorrhoeae. Gene. 1990 Sep 1;93(1):151–156. doi: 10.1016/0378-1119(90)90151-g. [DOI] [PubMed] [Google Scholar]
  11. Haas R., Meyer T. F. The repertoire of silent pilus genes in Neisseria gonorrhoeae: evidence for gene conversion. Cell. 1986 Jan 17;44(1):107–115. doi: 10.1016/0092-8674(86)90489-7. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Hagblom P., Segal E., Billyard E., So M. Intragenic recombination leads to pilus antigenic variation in Neisseria gonorrhoeae. Nature. 1985 May 9;315(6015):156–158. doi: 10.1038/315156a0. [DOI] [PubMed] [Google Scholar]
  14. Howell-Adams B., Wainwright L. A., Seifert H. S. The size and position of heterologous insertions in a silent locus differentially affect pilin recombination in Neisseria gonorrhoeae. Mol Microbiol. 1996 Nov;22(3):509–522. doi: 10.1046/j.1365-2958.1996.00128.x. [DOI] [PubMed] [Google Scholar]
  15. Jonsson A. B., Ilver D., Falk P., Pepose J., Normark S. Sequence changes in the pilus subunit lead to tropism variation of Neisseria gonorrhoeae to human tissue. Mol Microbiol. 1994 Aug;13(3):403–416. doi: 10.1111/j.1365-2958.1994.tb00435.x. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. KELLOGG D. S., Jr, PEACOCK W. L., Jr, DEACON W. E., BROWN L., PIRKLE D. I. NEISSERIA GONORRHOEAE. I. VIRULENCE GENETICALLY LINKED TO CLONAL VARIATION. J Bacteriol. 1963 Jun;85:1274–1279. doi: 10.1128/jb.85.6.1274-1279.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Koomey J. M., Falkow S. Cloning of the recA gene of Neisseria gonorrhoeae and construction of gonococcal recA mutants. J Bacteriol. 1987 Feb;169(2):790–795. doi: 10.1128/jb.169.2.790-795.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Koomey M., Gotschlich E. C., Robbins K., Bergström S., Swanson J. Effects of recA mutations on pilus antigenic variation and phase transitions in Neisseria gonorrhoeae. Genetics. 1987 Nov;117(3):391–398. doi: 10.1093/genetics/117.3.391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kowalczykowski S. C., Dixon D. A., Eggleston A. K., Lauder S. D., Rehrauer W. M. Biochemistry of homologous recombination in Escherichia coli. Microbiol Rev. 1994 Sep;58(3):401–465. doi: 10.1128/mr.58.3.401-465.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lin Y. P., Sharer J. D., March P. E. GTPase-dependent signaling in bacteria: characterization of a membrane-binding site for era in Escherichia coli. J Bacteriol. 1994 Jan;176(1):44–49. doi: 10.1128/jb.176.1.44-49.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Long C. D., Madraswala R. N., Seifert H. S. Comparisons between colony phase variation of Neisseria gonorrhoeae FA1090 and pilus, pilin, and S-pilin expression. Infect Immun. 1998 May;66(5):1918–1927. doi: 10.1128/iai.66.5.1918-1927.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. March P. E., Lerner C. G., Ahnn J., Cui X., Inouye M. The Escherichia coli Ras-like protein (Era) has GTPase activity and is essential for cell growth. Oncogene. 1988 Jun;2(6):539–544. [PubMed] [Google Scholar]
  24. March P. E. Membrane-associated GTPases in bacteria. Mol Microbiol. 1992 May;6(10):1253–1257. doi: 10.1111/j.1365-2958.1992.tb00845.x. [DOI] [PubMed] [Google Scholar]
  25. Mehr I. J., Seifert H. S. Random shuttle mutagenesis: gonococcal mutants deficient in pilin antigenic variation. Mol Microbiol. 1997 Mar;23(6):1121–1131. doi: 10.1046/j.1365-2958.1997.2971660.x. [DOI] [PubMed] [Google Scholar]
  26. Meier T. I., Peery R. B., Jaskunas S. R., Zhao G. 16S rRNA is bound to era of Streptococcus pneumoniae. J Bacteriol. 1999 Sep;181(17):5242–5249. doi: 10.1128/jb.181.17.5242-5249.1999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Morse S. A., Bartenstein L. Factors affecting autolysis of Neisseria gonorrhoeae. Proc Soc Exp Biol Med. 1974 Apr;145(4):1418–1421. doi: 10.3181/00379727-145-38025. [DOI] [PubMed] [Google Scholar]
  28. Pillutla R. C., Sharer J. D., Gulati P. S., Wu E., Yamashita Y., Lerner C. G., Inouye M., March P. E. Cross-species complementation of the indispensable Escherichia coli era gene highlights amino acid regions essential for activity. J Bacteriol. 1995 Apr;177(8):2194–2196. doi: 10.1128/jb.177.8.2194-2196.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Ryder L., Sharples G. J., Lloyd R. G. Recombination-dependent growth in exonuclease-depleted recBC sbcBC strains of Escherichia coli K-12. Genetics. 1996 Jul;143(3):1101–1114. doi: 10.1093/genetics/143.3.1101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Segal E., Billyard E., So M., Storzbach S., Meyer T. F. Role of chromosomal rearrangement in N. gonorrhoeae pilus phase variation. Cell. 1985 Feb;40(2):293–300. doi: 10.1016/0092-8674(85)90143-6. [DOI] [PubMed] [Google Scholar]
  32. Segal E., Hagblom P., Seifert H. S., So M. Antigenic variation of gonococcal pilus involves assembly of separated silent gene segments. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2177–2181. doi: 10.1073/pnas.83.7.2177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Seifert H. S., Chen E. Y., So M., Heffron F. Shuttle mutagenesis: a method of transposon mutagenesis for Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1986 Feb;83(3):735–739. doi: 10.1073/pnas.83.3.735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Seifert H. S., Wright C. J., Jerse A. E., Cohen M. S., Cannon J. G. Multiple gonococcal pilin antigenic variants are produced during experimental human infections. J Clin Invest. 1994 Jun;93(6):2744–2749. doi: 10.1172/JCI117290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. 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]
  37. Swanson J., Barrera O. Gonococcal pilus subunit size heterogeneity correlates with transitions in colony piliation phenotype, not with changes in colony opacity. J Exp Med. 1983 Nov 1;158(5):1459–1472. doi: 10.1084/jem.158.5.1459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Swanson J., Bergstrom S., Boslego J., Koomey M. Gene conversion accounts for pilin structural changes and for reversible piliation "phase" changes in gonococci. Antonie Van Leeuwenhoek. 1987;53(6):441–446. doi: 10.1007/BF00415500. [DOI] [PubMed] [Google Scholar]
  39. Swanson J., Bergström S., Robbins K., Barrera O., Corwin D., Koomey J. M. Gene conversion involving the pilin structural gene correlates with pilus+ in equilibrium with pilus- changes in Neisseria gonorrhoeae. Cell. 1986 Oct 24;47(2):267–276. doi: 10.1016/0092-8674(86)90449-6. [DOI] [PubMed] [Google Scholar]
  40. Swanson J., Kraus S. J., Gotschlich E. C. Studies on gonococcus infection. I. Pili and zones of adhesion: their relation to gonococcal growth patterns. J Exp Med. 1971 Oct 1;134(4):886–906. doi: 10.1084/jem.134.4.886. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Swanson J. Studies on gonococcus infection. IV. Pili: their role in attachment of gonococci to tissue culture cells. J Exp Med. 1973 Mar 1;137(3):571–589. doi: 10.1084/jem.137.3.571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Virji M., Everson J. S., Lambden P. R. Effect of anti-pilus antisera on virulence of variants of Neisseria gonorrhoeae for cultured epithelial cells. J Gen Microbiol. 1982 May;128(5):1095–1100. doi: 10.1099/00221287-128-5-1095. [DOI] [PubMed] [Google Scholar]
  43. Virji M., Heckels J. E. The role of common and type-specific pilus antigenic domains in adhesion and virulence of gonococci for human epithelial cells. J Gen Microbiol. 1984 May;130(5):1089–1095. doi: 10.1099/00221287-130-5-1089. [DOI] [PubMed] [Google Scholar]
  44. Virji M., Heckels J. E., Watt P. J. Monoclonal antibodies to gonococcal pili: studies on antigenic determinants on pili from variants of strain P9. J Gen Microbiol. 1983 Jun;129(6):1965–1973. doi: 10.1099/00221287-129-6-1965. [DOI] [PubMed] [Google Scholar]
  45. Wainwright L. A., Frangipane J. V., Seifert H. S. Analysis of protein binding to the Sma/Cla DNA repeat in pathogenic Neisseriae. Nucleic Acids Res. 1997 Apr 1;25(7):1362–1368. doi: 10.1093/nar/25.7.1362. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wainwright L. A., Pritchard K. H., Seifert H. S. A conserved DNA sequence is required for efficient gonococcal pilin antigenic variation. Mol Microbiol. 1994 Jul;13(1):75–87. doi: 10.1111/j.1365-2958.1994.tb00403.x. [DOI] [PubMed] [Google Scholar]
  47. West A. B., Roberts T. M., Kolodner R. D. Regulation of the reverse transcriptase of human immunodeficiency virus type 1 by dNTPs. Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9720–9724. doi: 10.1073/pnas.89.20.9720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Wright C. J., Jerse A. E., Cohen M. S., Cannon J. G., Seifert H. S. Nonrepresentative PCR amplification of variable gene sequences in clinical specimens containing dilute, complex mixtures of microorganisms. J Clin Microbiol. 1994 Feb;32(2):464–468. doi: 10.1128/jcm.32.2.464-468.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES