Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 May;177(9):2475–2480. doi: 10.1128/jb.177.9.2475-2480.1995

Variable expression of class 1 outer membrane protein in Neisseria meningitidis is caused by variation in the spacing between the -10 and -35 regions of the promoter.

A van der Ende 1, C T Hopman 1, S Zaat 1, B B Essink 1, B Berkhout 1, J Dankert 1
PMCID: PMC176907  PMID: 7730280

Abstract

The class 1 outer membrane protein encoded by the porA gene of Neisseria meningitidis is a candidate for a vaccine against meningococcal infection. The expression of class 1 outer membrane protein displays phase variation between three expression levels. Northern (RNA) blot and primer extension analysis revealed that this phase variation is regulated at the transcriptional level. The start site for transcription is located 59 bp upstream of the translational initiation codon. Sequence analysis of the promoter region of the porA gene of a variant without class 1 protein expression revealed nine contiguous guanidine residues between the -10 and -35 domains. Comparison of promoter sequences of different phase variants indicated that the length of the polyguanidine stretch correlated with the expression level of the class 1 outer membrane protein; the presence of 11, 10, or 9 contiguous guanidine residues results in high levels, medium levels, or no expression of class 1 mRNA, respectively. These results suggest that the variable porA expression levels seen in different isolates are modulated by guanidine residue insertion and/or deletion due to slipped-strand mispairing on the polyguanidine stretch within the intervening sequence of the -35 and -10 regions of the promoter. The phase variation of class 1 outer membrane protein may provide a molecular mechanism to evade the host immune defense. Therefore, the protective efficacy of a vaccine based on class 1 outer membrane protein may be questioned.

Full Text

The Full Text of this article is available as a PDF (266.7 KB).

Selected References

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

  1. Barlow A. K., Heckels J. E., Clarke I. N. The class 1 outer membrane protein of Neisseria meningitidis: gene sequence and structural and immunological similarities to gonococcal porins. Mol Microbiol. 1989 Feb;3(2):131–139. doi: 10.1111/j.1365-2958.1989.tb01802.x. [DOI] [PubMed] [Google Scholar]
  2. Berkhout B., Gatignol A., Silver J., Jeang K. T. Efficient trans-activation by the HIV-2 Tat protein requires a duplicated TAR RNA structure. Nucleic Acids Res. 1990 Apr 11;18(7):1839–1846. doi: 10.1093/nar/18.7.1839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Crowe B. A., Wall R. A., Kusecek B., Neumann B., Olyhoek T., Abdillahi H., Hassan-King M., Greenwood B. M., Poolman J. T., Achtman M. Clonal and variable properties of Neisseria meningitidis isolated from cases and carriers during and after an epidemic in The Gambia, West Africa. J Infect Dis. 1989 Apr;159(4):686–700. doi: 10.1093/infdis/159.4.686. [DOI] [PubMed] [Google Scholar]
  4. Dryden S. C., Kaplan S. Identification of cis-acting regulatory regions upstream of the rRNA operons of Rhodobacter sphaeroides. J Bacteriol. 1993 Oct;175(20):6392–6402. doi: 10.1128/jb.175.20.6392-6402.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Frasch C. E., Zollinger W. D., Poolman J. T. Serotype antigens of Neisseria meningitidis and a proposed scheme for designation of serotypes. Rev Infect Dis. 1985 Jul-Aug;7(4):504–510. doi: 10.1093/clinids/7.4.504. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Harley C. B., Reynolds R. P. Analysis of E. coli promoter sequences. Nucleic Acids Res. 1987 Mar 11;15(5):2343–2361. doi: 10.1093/nar/15.5.2343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Higgins D. G., Sharp P. M. CLUSTAL: a package for performing multiple sequence alignment on a microcomputer. Gene. 1988 Dec 15;73(1):237–244. doi: 10.1016/0378-1119(88)90330-7. [DOI] [PubMed] [Google Scholar]
  9. Hitchcock P. J. Unified nomenclature for pathogenic Neisseria species. Clin Microbiol Rev. 1989 Apr;2 (Suppl):S64–S65. doi: 10.1128/cmr.2.suppl.s64. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Josaitis C. A., Gaal T., Ross W., Gourse R. L. Sequences upstream of the-35 hexamer of rrnB P1 affect promoter strength and upstream activation. Biochim Biophys Acta. 1990 Aug 27;1050(1-3):307–311. doi: 10.1016/0167-4781(90)90186-6. [DOI] [PubMed] [Google Scholar]
  11. Koo H. S., Wu H. M., Crothers D. M. DNA bending at adenine . thymine tracts. Nature. 1986 Apr 10;320(6062):501–506. doi: 10.1038/320501a0. [DOI] [PubMed] [Google Scholar]
  12. Langenberg W., Rauws E. A., Widjojokusumo A., Tytgat G. N., Zanen H. C. Identification of Campylobacter pyloridis isolates by restriction endonuclease DNA analysis. J Clin Microbiol. 1986 Sep;24(3):414–417. doi: 10.1128/jcm.24.3.414-417.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Maiden M. C., Suker J., McKenna A. J., Bygraves J. A., Feavers I. M. Comparison of the class 1 outer membrane proteins of eight serological reference strains of Neisseria meningitidis. Mol Microbiol. 1991 Mar;5(3):727–736. doi: 10.1111/j.1365-2958.1991.tb00743.x. [DOI] [PubMed] [Google Scholar]
  14. McGuinness B., Barlow A. K., Clarke I. N., Farley J. E., Anilionis A., Poolman J. T., Heckels J. E. Deduced amino acid sequences of class 1 protein (PorA) from three strains of Neisseria meningitidis. Synthetic peptides define the epitopes responsible for serosubtype specificity. J Exp Med. 1990 Jun 1;171(6):1871–1882. doi: 10.1084/jem.171.6.1871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Meyer T. F., Gibbs C. P., Haas R. Variation and control of protein expression in Neisseria. Annu Rev Microbiol. 1990;44:451–477. doi: 10.1146/annurev.mi.44.100190.002315. [DOI] [PubMed] [Google Scholar]
  16. Munkley A., Tinsley C. R., Virji M., Heckels J. E. Blocking of bactericidal killing of Neisseria meningitidis by antibodies directed against class 4 outer membrane protein. Microb Pathog. 1991 Dec;11(6):447–452. doi: 10.1016/0882-4010(91)90041-8. [DOI] [PubMed] [Google Scholar]
  17. Parker J. D., Rabinovitch P. S., Burmer G. C. Targeted gene walking polymerase chain reaction. Nucleic Acids Res. 1991 Jun 11;19(11):3055–3060. doi: 10.1093/nar/19.11.3055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Plaskon R. R., Wartell R. M. Sequence distributions associated with DNA curvature are found upstream of strong E. coli promoters. Nucleic Acids Res. 1987 Jan 26;15(2):785–796. doi: 10.1093/nar/15.2.785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Poolman J. T., de Marie S., Zanen H. C. Variability of low-molecular-weight, heat-modifiable outer membrane proteins of Neisseria meningitidis. Infect Immun. 1980 Dec;30(3):642–648. doi: 10.1128/iai.30.3.642-648.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Robertson B. D., Meyer T. F. Genetic variation in pathogenic bacteria. Trends Genet. 1992 Dec;8(12):422–427. doi: 10.1016/0168-9525(92)90325-x. [DOI] [PubMed] [Google Scholar]
  21. Sarkari J., Pandit N., Moxon E. R., Achtman M. Variable expression of the Opc outer membrane protein in Neisseria meningitidis is caused by size variation of a promoter containing poly-cytidine. Mol Microbiol. 1994 Jul;13(2):207–217. doi: 10.1111/j.1365-2958.1994.tb00416.x. [DOI] [PubMed] [Google Scholar]
  22. Saukkonen K., Abdillahi H., Poolman J. T., Leinonen M. Protective efficacy of monoclonal antibodies to class 1 and class 3 outer membrane proteins of Neisseria meningitidis B:15:P1.16 in infant rat infection model: new prospects for vaccine development. Microb Pathog. 1987 Oct;3(4):261–267. doi: 10.1016/0882-4010(87)90059-3. [DOI] [PubMed] [Google Scholar]
  23. Saukkonen K., Leinonen M., Abdillahi H., Poolman J. T. Comparative evaluation of potential components for group B meningococcal vaccine by passive protection in the infant rat and in vitro bactericidal assay. Vaccine. 1989 Aug;7(4):325–328. doi: 10.1016/0264-410x(89)90194-1. [DOI] [PubMed] [Google Scholar]
  24. Seifert H. S., Ajioka R., So M. Alternative model for Neisseria gonorrhoeae pilin variation. Vaccine. 1988 Apr;6(2):107–109. doi: 10.1016/s0264-410x(88)80009-4. [DOI] [PubMed] [Google Scholar]
  25. Siebenlist U., Simpson R. B., Gilbert W. E. coli RNA polymerase interacts homologously with two different promoters. Cell. 1980 Jun;20(2):269–281. doi: 10.1016/0092-8674(80)90613-3. [DOI] [PubMed] [Google Scholar]
  26. Stephens D. S., McGee Z. A. Association of virulence of Neisseria meningitidis with transparent colony type and low-molecular-weight outer membrane proteins. J Infect Dis. 1983 Feb;147(2):282–292. doi: 10.1093/infdis/147.2.282. [DOI] [PubMed] [Google Scholar]
  27. Tinsley C. R., Heckels J. E. Variation in the expression of pili and outer membrane protein by Neisseria meningitidis during the course of meningococcal infection. J Gen Microbiol. 1986 Sep;132(9):2483–2490. doi: 10.1099/00221287-132-9-2483. [DOI] [PubMed] [Google Scholar]
  28. Tommassen J., Vermeij P., Struyvé M., Benz R., Poolman J. T. Isolation of Neisseria meningitidis mutants deficient in class 1 (porA) and class 3 (porB) outer membrane proteins. Infect Immun. 1990 May;58(5):1355–1359. doi: 10.1128/iai.58.5.1355-1359.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tsai C. M., Frasch C. E., Mocca L. F. Five structural classes of major outer membrane proteins in Neisseria meningitidis. J Bacteriol. 1981 Apr;146(1):69–78. doi: 10.1128/jb.146.1.69-78.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Willems R., Paul A., van der Heide H. G., ter Avest A. R., Mooi F. R. Fimbrial phase variation in Bordetella pertussis: a novel mechanism for transcriptional regulation. EMBO J. 1990 Sep;9(9):2803–2809. doi: 10.1002/j.1460-2075.1990.tb07468.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Woods J. P., Cannon J. G. Variation in expression of class 1 and class 5 outer membrane proteins during nasopharyngeal carriage of Neisseria meningitidis. Infect Immun. 1990 Feb;58(2):569–572. doi: 10.1128/iai.58.2.569-572.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yogev D., Rosengarten R., Watson-McKown R., Wise K. S. Molecular basis of Mycoplasma surface antigenic variation: a novel set of divergent genes undergo spontaneous mutation of periodic coding regions and 5' regulatory sequences. EMBO J. 1991 Dec;10(13):4069–4079. doi: 10.1002/j.1460-2075.1991.tb04983.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. van Ham S. M., van Alphen L., Mooi F. R., van Putten J. P. Phase variation of H. influenzae fimbriae: transcriptional control of two divergent genes through a variable combined promoter region. Cell. 1993 Jun 18;73(6):1187–1196. doi: 10.1016/0092-8674(93)90647-9. [DOI] [PubMed] [Google Scholar]
  34. van der Ley P., van der Biezen J., Hohenstein P., Peeters C., Poolman J. T. Use of transformation to construct antigenic hybrids of the class 1 outer membrane protein in Neisseria meningitidis. Infect Immun. 1993 Oct;61(10):4217–4224. doi: 10.1128/iai.61.10.4217-4224.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES