1887

Abstract

A DNA transformation dose-response curve of piliated (P) gonococci with the use of cloned DNA containing a fusion showed saturation at high and low levels of transforming DNA. At low DNA concentrations, transformation of the P strain MS11-A was effectively inhibited by a 1000-fold molar excess of the gonococcal transformation uptake sequence (GCUS). The same molar excess of the GCUS did not inhibit transformation of MS11-A at high DNA concentrations. In MS11-B2, a nonpiliated (P), pilin-nonproducing, isogenic variant of MS11-A, the GCUS did not inhibit transformation at any level of transforming DNA. These data suggest that two mechanisms of transformation exist in P cells: one which utilizes the GCUS and one which does not. In MS11-B2 P cells, no evidence was found for the presence of the GCUS-dependent mechanism, suggesting that transformation in this background occurs solely by the GCUS-independent mechanism.

Keyword(s): competition , dose-response and pili
Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-142-10-2839
1996-10-01
2021-05-18
Loading full text...

Full text loading...

/deliver/fulltext/micro/142/10/mic-142-10-2839.html?itemId=/content/journal/micro/10.1099/13500872-142-10-2839&mimeType=html&fmt=ahah

References

  1. Birnboim H. C., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523
    [Google Scholar]
  2. Biswas G. D., Sox T., Blackman E., Sparling P. F. 1977; Factors affecting genetic transformation of Neisseria gonorrhoeae . J Bacteriol 129:983–992
    [Google Scholar]
  3. Biswas G. D., Lacks S. A., Sparling P. F. 1989; Transformation-deficient mutants of piliated Neisseria gonorrhoeae . J Bacteriol 171:657–664
    [Google Scholar]
  4. Boyle-Vavra S., Seifert H. S. 1993; Shuttle mutagenesis: two mini-transposons for gene mapping and for lacZ transcriptional fusions in Neisseria gonorrhoeae . Gene 129:51–57
    [Google Scholar]
  5. Burnstein K. L., Dyer D. W., Sparling P. F. 1988; Preferential uptake of restriction fragments from a gonococcal cryptic plasmid by competent Neisseria gonorrhoeae . J Gen Microbiol 134:547–557
    [Google Scholar]
  6. Cannon J. G., Sparling P. F. 1984; The genetics of the gonococcus. Annu Rev Microbiol 38:111–133
    [Google Scholar]
  7. Catlin B. W., Cunningham L. S. 1961; Transforming activities and base contents of deoxyribonucleate preparations from various neisseriae. J Gen Microbiol 26:303–312
    [Google Scholar]
  8. Clewell D. B., Helinski D. R. 1969; Supercoiled circular DNA- protein complex in Escherichia coli: purification and induced conversion to an open circular form. Biochemistry 9:4428–4440
    [Google Scholar]
  9. Close T. J., Rodriguez R. L. 1982; Construction and characterization of the chloramphenicol-resistance gene cartridge: a new approach to the transcriptional mapping of extrachromosomal elements. Gene 20:305–316
    [Google Scholar]
  10. Dorward D. W., Garon C. F. 1989; DNA-binding proteins in cells and membrane blebs of Neisseria gonorrhoeae . J Bacteriol 171:4196–4201
    [Google Scholar]
  11. Dougherty T. J., Asmus A., Tomasz A. 1979; Specificity of DNA uptake in genetic transformation of gonococci. Biochem Biophys Res Commun 86:97–104
    [Google Scholar]
  12. Elkins C., Thomas C. E., Seifert H. S., Sparling P. F. 1991; Species-specific uptake of DNA by gonococci is mediated by a 10- base-pair sequence. J Bacteriol 173:3911–3913
    [Google Scholar]
  13. Facius D., Meyer T. F. 1993; A novel determinant (comA )essential for natural transformation competence in Neisseria gonorrhoeae and the effect of a com A mutation on pilin variation. Mol Microbiol 10:699–712
    [Google Scholar]
  14. Feinberg A. P., Vogelstein B. 1983; A technique for radio-labeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
    [Google Scholar]
  15. Frosch M., Meyer T. F. 1992; Transformation-mediated exchange of virulence determinants by co-cultivation of pathogenic Neisseriae . FEMS Microbiol Lett 79:345–349
    [Google Scholar]
  16. Gibbs C. P., Reimann B. Y., Schultz E., Kaufmann A., Haas R., Meyer T. F. 1989; Reassortment of pilin genes in Neisseria gonorrhoeae occurs by two distinct mechanisms. Nature 338:651–652
    [Google Scholar]
  17. Goodman S. D., Scocca J. J. 1988; Identification and arrangement of the DNA sequence recognized in specific transformation of Neisseria gonorrhoeae . Proc Natl Acad Sci USA 856982–6986
    [Google Scholar]
  18. Haas R., Meyer T. F. 1986; The repertoire of silent pilus genes in Neisseria gonorrhoeae: evidence for gene conversion. Cell 44:107–115
    [Google Scholar]
  19. Haas R., Veit S., Meyer T. F. 1992; Silent pilin genes of Neisseria gonorrhoeae MS11 and the occurrence of related hyper-variant sequences among other gonococcal isolates. Mol Microbiol 6:197–208
    [Google Scholar]
  20. Halter R., Pohlner J., Meyer T. F. 1989; Mosaic-like organization of IgA protease genes in Neisseria gonorrhoeae generated by horizontal genetic exchange in vivo. EMBO J 8:2737–2744
    [Google Scholar]
  21. Hebeler B. H., Young F. E. 1975; Autolysis of Neisseria gonorrhoeae . J Bacteriol 122:385–392
    [Google Scholar]
  22. Hobbs M., Mattick J. S. 1993; 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 10:233–243
    [Google Scholar]
  23. Joenje H., Konings W. N., Venema G. 1975; Interactions between exogenous deoxyribonucleic acid and membrane vesicles isolated from competent and noncompetent Bacillus subtilis . J Bacteriol 121:771–776
    [Google Scholar]
  24. Jonsson A. B., Nyberg G., Normark S. 1991; Phase variation of gonococcal pili by frameshift mutation in pilC, a novel gene for pilus assembly. EMBO J 10:477–488
    [Google Scholar]
  25. Kellogg D. S. Jr Peacock W. L., Deacon W. E., Brown L. 1963; Neisseria gonorrhoeae. I. Virulence genetically linked to clonal variation. J Bacteriol 85:1274–1279
    [Google Scholar]
  26. Koomey J. M., Falkow S. 1987; Cloning of the recA gene of Neisseria gonorrhoeae and construction of gonococcal recA mutants. J Bacteriol 169:790–795
    [Google Scholar]
  27. Koomey M., Fox R., Brossay L., Hebert J. 1994; The gonococcal pilT protein plays an essential role in pilus-associated phenotypes of twitching motility and natural competence for transformation. In Proceedings of the 9th International Pathogenesis Neisseria Conference Winchester, UK:
    [Google Scholar]
  28. Mathis LS., Scocca J. J. 1984; On the role of pili in transformation of Neisseria gonorrhoeae . J Gen Microbiol 130:3165–3173
    [Google Scholar]
  29. Meyer T. F., Billyard E., Haas R., Storzbach S., So M. 1984; Pilus genes of Neisseria gonorrhoeae: chromosomal organization and DNA sequence. Proc Natl Acad Sci USA 81:6110–6114
    [Google Scholar]
  30. Norlander L., Davies J., Norqvist A., Normark S. 1979; Genetic basis for colonial variation in Neisseria gonorrhoeae . J Bacteriol 138:762–769
    [Google Scholar]
  31. Rudel T., Facius D., Barten R., Scheuerpflug I., Nonnenmacher E. , Meyer T. F. 1995a; Role of pili and the phase variable PilC protein in natural competence for transformation of Neisseria gonorrhoeae . Proc Natl Acad Sci USA 92:7986–7990
    [Google Scholar]
  32. Rudel T., Scheuerpflug I., Meyer T. F. 1995b; Neisseria PilC protein identified as type-4 pilus tip-located adhesin. Nature 373:357–359
    [Google Scholar]
  33. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  34. Schwalbe R. S., Cannon J. G. 1986; Genetic transformation of genes for protein II in Neisseria gonorrhoeae . J Bacteriol 167:186–190
    [Google Scholar]
  35. Segal E., Hagblom P., Seifert H. S., So M. 1986; Antigenic variation of gonococcal pilus involves assembly of separated silent gene segments. Proc Natl Acad Sci USA 832177–2181
    [Google Scholar]
  36. Seifert H. S. 1992; Molecular mechanisms of antigenic variation in Neisseria gonorrhoeae . In Molecular and Cell Biology of Sexually Transmitted Diseases pp. 1–22 Edited by Wright D., Archard L. London: Chapman & Hall;
    [Google Scholar]
  37. Seifert H. S., So M. 1991; Genetic systems in pathogenic Neisseriae . Methods Engymol 204:342–357
    [Google Scholar]
  38. Seifert H. S., Ajioka R. S., Marchal C., Sparling P. F., So M. 1988; DNA transformation leads to pilin antigenic variation in Neisseria gonorrhoeae . Nature 336:392–395
    [Google Scholar]
  39. Seifert H. S., Ajioka R. S., Paruchuri D., Heffron F., So M. 1990; Shuttle mutagenesis of Neisseria gonorrhoeae: pilin null mutations lower DNA transformation competence. J Bacteriol 172:40–46
    [Google Scholar]
  40. Sparling P. F. 1966; Genetic transformation of Neisseria gonorrhoeae to streptomycin resistance. J Bacteriol 92:1364–1371
    [Google Scholar]
  41. Sparling P. F., Biswas G. D., Sox T. E. 1977; Transformation of the gonococcus. In The Gonococcus pp. 177–198 Edited by Roberts R. B. New York: John Wiley;
    [Google Scholar]
  42. Spratt B. G., Bowler L. D., Zhang Q. Y., Zhou J., Smith J. M. 1992; Role of interspecies transfer of chromosomal genes in the evolution of penicillin resistance in pathogenic and commensal Neisseria species. J Mol Evol 34:115–125
    [Google Scholar]
  43. Stein D. C. 1991; Transformation of Neisseria gonorrhoeae: physical requirements of the transforming DNA. Can J Microbiol 37:345–349
    [Google Scholar]
  44. Swanson J., Bergstrom S., Barrera O., Robbins K., Corwin D. 1985; Pilus gonococcal variants. Evidence for multiple forms of piliation control. J Exp Med 162:729–744
    [Google Scholar]
  45. Swanson J., Morrison S., Barrera O., Hill S. 1990; Piliation changes in transformation-defective gonococci. J Exp Med 171:2131–2139
    [Google Scholar]
  46. Taha M. K., So M., Seifert H. S., Billyard E., Marchal C. 1988; Pilin expression in Neisseria gonorrhoeae is under both positive and negative transcriptional control. EMBO J 7:4367–4378
    [Google Scholar]
  47. Zhang Q. Y., DeRyckere D., Lauer P., Koomey M. 1992; Gene conversion in Neisseria gonorrhoeae: evidence for its role in pilus antigenic variation. Proc Natl Acad Sci USA 895366–5370
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-142-10-2839
Loading
/content/journal/micro/10.1099/13500872-142-10-2839
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error