A real-time semi-quantitative RT-PCR assay demonstrates that the pilE sequence dictates the frequency and characteristics of pilin antigenic variation in Neisseria gonorrhoeae

Nucleic Acids Research

Volumn 33, Issue 10, 2005, Pages 3363-3371

  Rohrer, Melissa S ^a   Lazio, Matthew P ^a,b   Seifert, H Steven ^a  

^a NORTHWESTERN UNIVERSITY   (United States)

^b UNIVERSITY OF IOWA   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

PILIN; FIMBRIA PROTEIN; PILE PROTEIN, NEISSERIA GONORRHOEAE;

ACCURACY; ANTIGENIC VARIATION; ARTICLE; BACTERIAL STRAIN; DNA SEQUENCE; GENE AMPLIFICATION; GENE IDENTIFICATION; GENETIC VARIABILITY; METHODOLOGY; NEISSERIA GONORRHOEAE; NONHUMAN; PILE GENE; PRIORITY JOURNAL; QUANTITATIVE ANALYSIS; REVERSE TRANSCRIPTION; REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION; RNA EXTRACTION; SILENT GENE; EVALUATION; GENE SILENCING; GENETICS; TIME;

NEISSERIA GONORRHOEAE;

ANTIGENIC VARIATION; FIMBRIAE PROTEINS; GENE SILENCING; NEISSERIA GONORRHOEAE; REVERSE TRANSCRIPTASE POLYMERASE CHAIN REACTION; SEQUENCE ANALYSIS, DNA; TIME FACTORS;

EID: 20444439947     PISSN: 03051048     EISSN: None     Source Type: Journal    
DOI: 10.1093/nar/gki650     Document Type: Article

References (34)

1. Virji,M. and Heckels,J.E. (1983) Antigenic cross-reactivity of Neisseria pili: Investigations with type- and species-specific monoclonal antibodies. J. Gen. Microbiol., 129, 2761-2768.
2. Zak,K., Diaz,J.-L., Jackson,D. and Heckels,J.E. (1984) Antigenic variation during infection with Neisseria gonorrhoeae: detection of antibodies to surface proteins in sera of patients with gonorrhea. J. Infect. Dis., 149, 166-174.
3. Brinton,C.C., Bryan,J., Dillon,J.-A., Guernia,N., Jackobson,L.J., Labik,A., Lee,S., Levine,A., Lim,S., McMichael,J. et al. (1978) In Brooks,G.F., Gotschlich,E.C., Holmes,K.K., Sawyer,W.D. and Young,F.E. (eds), Immunobiology of Neisseria gonorrhoeae. American Society for Microbiology, Washington, DC, pp. 155-178.
4. Rudel,T., Boxberger,H.J. and Meyer,T.F. (1995) Pilus biogenesis and epithelial cell adherence of Neisseria gonorrhoeae pilC double knock-out mutants. Mol. Microbiol., 17, 1057-1071.
5. Nassif,X. and So,M. (1995) Interaction of pathogenic Neisseriae with nonphagocytic cells. Clin. Microbiol. Rev., 8, 376-388.
6. Sparling,P.F. (1966) Genetic transformation of Neisseria gonorrhoeae to streptomycin resistance. J. Bacteriol., 92, 1364-1371.
7. Seifert,H.S., Ajioka,R.S., Paruchuri,D., Heffron,F. and So,M. (1990) Shuttle mutagenesis of Neisseria gonorrhoeae: pilin null mutations lower DNA transformation competence. J. Bacteriol., 172, 40-46.
8. Freitag,N.E., Seifert,H.S. and Koomey,M. (1995) Characterization of the pilF-pilD pilus-assembly locus of Neisseria gonorrhoeae. Mol. Microbiol., 95, 575-586.
9. Hamrick,T.S., Dempsey,J.A., Cohen,M.S. and Cannon,J.G. (2001) Antigenic variation of gonococcal pilin expression in vivo: Analysis of the strain FA1090 pilin repertoire and identification of the pilS gene copies recombining with pilE during experimental human infection. Microbiology, 147, 839-849.
10. Hagblom,P., Segal,E., Billyard,E. and So,M. (1985) Intragenic recombination leads to pilus antigenic variation in Neisseria gonorrhoeae. Nature, 315, 156-158.
11. Meyer,T.F., Mlawer,N. and So,M. (1982) Pilus expression in Neisseria gonorrhoeae involves chromosomal rearrangement. Cell, 30, 45-52.
12. Segal,E., Billyard,E., So,M., Storzbach,S. and Meyer,T.F. (1985) Role of chromosomal rearrangement in N.gonorrhoeae pilus phase variation. Cell, 40, 293-300.
13. Haas,R. and Meyer,T.F. (1986) The repertoire of silent pilus genes in Neisseria gonorrhoeae: Evidence for gene conversion. Cell, 44, 107-115.
14. Segal,E., Hagblom,P., Seifert,H.S. and So,M. (1986) Antigenic variation of gonococcal pilus involves assembly of separated silent gene segments. Proc. Natl Acad. Sci. USA, 83, 2177-2181.
15. Howell-Adams,B., Wainwright,L.A. and Seifert,H.S. (1996) The size and position of heterologous insertions in a silent locus differentially affect pilin recombination in Neisseria gonorrhoeae. Mol. Microbiol. 22, 509-522.
16. Howell-Adams,B. and Seifert,H.S. (1999) Insertion mutations in pilE differentially alter gonococcal pilin antigenic variation. J. Bacteriol., 181, 6133-6141.
17. Wainwright,L.A., Pritchard,K.H. and Seifert,H.S. (1994) A conserved DNA sequence is required for efficient gonococcal pilin antigenic variation. Mol. Microbiol., 13, 75-87.
18. Mehr,I.J. and Seifert,H.S. (1998) Differential roles of homologous recombination pathways in Neisseria gonorrhoeae pilin antigenic variation, DNA transformation, and DNA repair. Mol. Microbiol., 30, 697-710.
19. Skaar,E.P., Lazio,M.P. and Seifert,H.S. (2002) Roles of the recJ and recN genes in homologous recombination and DNA repair pathways of Neisseria gonorrhoeae. J. Bacteriol., 184, 919-927.
20. Sechman,E.V., Rohrer,M.S. and Seifert,H.S. (2005) A genetic screen identifies genes important for iron-dependent and iron-independent pilin antigenic variation in Neisseria gonorrhoeae. Mol. Microbiol., in press.
21. Kline,K.A. and Seifert,H.S. (2005) Role of the Rep helicase gene in homologous recombination in Neisseria gonorrhoeae. J. Bacteriol., 187, 2903-2907.
22. Stohl,E.A. and Seifert,H.S. (2001) The recX gene potentiates homologous recombination in Neisseria gonorrhoeae. Mol. Microbiol. 40, 1301-1310.
23. Kline,K.A. and Seifert,H.S. (2005) Mutation of the PriA gene of Neisseria gonorrhoeae affects DNA transformation and DNA repair. J. Bacteriol., in press.
24. Serkin,C.D. and Seifert,H.S. (1998) Frequency of pilin antigenic variation in Neisseria gonorrhoeae. J. Bacteriol., 180, 1955-1958.
25. Stohl,E.A., Brockman,J.P., Burkle,K.L., Morimatsu,K., Kowalczykowski,S.C. and Seifert,H.S. (2003) Escherichia coli RecX inhibits RecA recombinase and coprotease activities in vitro and in vivo. J. Biol. Chem., 278, 2278-2285.
26. Okeke,C.N., Tsuboi,R. and Ogawa,H. (2001) Quantification of Candida albicans actin mRNA by the LightCycler system as a means of assessing viability in a model of cutaneous candidiasis. J. Clin. Microbiol. 39, 3491-3494.
27. Seifert,H.S., Wright,C.J., Jerse,A.E., Cohen,M.S. and Cannon,J.G. (1994) Multiple gonococcal pilin antigenic variants are produced during experimental human infections. J. Clin. Invest., 93, 2744-2749.
28. Seifert,H.S. (1997) Insertionally inactivated and inducible recA alleles for use in Neisseria. Gene, 188, 215-220.
29. Kellogg,D.S.,Jr, Peacock,W.L., Deacon,W.E., Brown,L. and Pirkle,C.I. (1963) Neisseria gonorrhoeae. I. Virulence genetically linked to clonial variation. J. Bacteriol., 85, 1274-1279.
30. Serkin,C.D. and Seifert,H.S. (2000) Iron availability regulates DNA recombination in Neisseria gonorrhoeae. Mol. Microbiol., 37, 1075-1086.
31. Meuer,S.C., Wittwer,C. and Nakagawara,K. (2001) Rapid Cycle Real-time PCR: Methods and Applications. Springer, Berlin, New York.
32. Wright,C.J., Jerse,A.E., Cohen,M.S., Cannon,J.G. and Seifert,H.S. (1994) Nonrepresentative PCR amplification of variable gene sequences in clinical specimens containing dilute, complex mixtures of microorganisms. J. Clin. Microbiol., 32, 464-468.
33. Sambrook,J., Fritsch,E.F. and Maniatis,T. (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
34. Koomey,M., Gotschlich,E.C., Robbins,K., Bergstrom,S. and Swanson,J. (1987) Effects of recA mutations on pilus antigenic variation and phase transitions in Neisseria gonorrhoeae. Genetics, 117, 391-398.