Abstract
The Campylobacter fetus surface-layer (S-layer) proteins mediate both complement resistance and antigenic variation in mammalian hosts. Wild-type strain 23D possesses the sapA gene, which encodes a 97-kDa S-layer protein, and several sapA homologs are present in both wild-type and mutant strains. Here we report that a cloned silent gene (sapA1) in C. fetus can express a functional full-length S-layer protein in Escherichia coli. Analysis of sapA and sapA1 and partial analysis of sapA2 indicate that a block of approximately 600 bp beginning upstream and continuing into the open reading frames is completely conserved, and then the sequences diverge completely, but immediately downstream of each gene is another conserved 50-bp sequence. Conservation of sapA1 among strains, the presence of a putative Chi (RecBCD recognition) site upstream of sapA, sapA1, and sapA2, and the sequence identities of the sapA genes suggest a system for homologous recombination. Comparison of the wild-type strain (23D) with a phenotypic variant (23D-11) indicates that variation is associated with removal of the divergent region of sapA from the expression locus and exchange with a corresponding region from a sapA homolog. We propose that site-specific reciprocal recombination between sapA homologs leads to expression of divergent S-layer proteins as one of the mechanisms that C. fetus uses for antigenic variation.
Full text
PDF![7265](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5242/47117/fc54783e6394/pnas01472-0380.png)
![7266](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5242/47117/2ece4d50b579/pnas01472-0381.png)
![7267](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5242/47117/d3db1493278e/pnas01472-0382.png)
![7268](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5242/47117/925f0663bb1c/pnas01472-0383.png)
![7269](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5242/47117/934c41e096f6/pnas01472-0384.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Abraham J. M., Freitag C. S., Clements J. R., Eisenstein B. I. An invertible element of DNA controls phase variation of type 1 fimbriae of Escherichia coli. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5724–5727. doi: 10.1073/pnas.82.17.5724. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Blake M. S., Johnston K. H., Russell-Jones G. J., Gotschlich E. C. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. doi: 10.1016/0003-2697(84)90320-8. [DOI] [PubMed] [Google Scholar]
- Blaser M. J., Gotschlich E. C. Surface array protein of Campylobacter fetus. Cloning and gene structure. J Biol Chem. 1990 Aug 25;265(24):14529–14535. [PubMed] [Google Scholar]
- Blaser M. J., Pei Z. Pathogenesis of Campylobacter fetus infections: critical role of high-molecular-weight S-layer proteins in virulence. J Infect Dis. 1993 Feb;167(2):372–377. doi: 10.1093/infdis/167.2.372. [DOI] [PubMed] [Google Scholar]
- Blaser M. J., Smith P. F., Hopkins J. A., Heinzer I., Bryner J. H., Wang W. L. Pathogenesis of Campylobacter fetus infections: serum resistance associated with high-molecular-weight surface proteins. J Infect Dis. 1987 Apr;155(4):696–706. doi: 10.1093/infdis/155.4.696. [DOI] [PubMed] [Google Scholar]
- Blaser M. J., Smith P. F., Repine J. E., Joiner K. A. Pathogenesis of Campylobacter fetus infections. Failure of encapsulated Campylobacter fetus to bind C3b explains serum and phagocytosis resistance. J Clin Invest. 1988 May;81(5):1434–1444. doi: 10.1172/JCI113474. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Borst P., Greaves D. R. Programmed gene rearrangements altering gene expression. Science. 1987 Feb 6;235(4789):658–667. doi: 10.1126/science.3544215. [DOI] [PubMed] [Google Scholar]
- Connell T. D., Black W. J., Kawula T. H., Barritt D. S., Dempsey J. A., Kverneland K., Jr, Stephenson A., Schepart B. S., Murphy G. L., Cannon J. G. Recombination among protein II genes of Neisseria gonorrhoeae generates new coding sequences and increases structural variability in the protein II family. Mol Microbiol. 1988 Mar;2(2):227–236. doi: 10.1111/j.1365-2958.1988.tb00024.x. [DOI] [PubMed] [Google Scholar]
- Dubreuil J. D., Kostrzynska M., Austin J. W., Trust T. J. Antigenic differences among Campylobacter fetus S-layer proteins. J Bacteriol. 1990 Sep;172(9):5035–5043. doi: 10.1128/jb.172.9.5035-5043.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dubreuil J. D., Logan S. M., Cubbage S., Eidhin D. N., McCubbin W. D., Kay C. M., Beveridge T. J., Ferris F. G., Trust T. J. Structural and biochemical analyses of a surface array protein of Campylobacter fetus. J Bacteriol. 1988 Sep;170(9):4165–4173. doi: 10.1128/jb.170.9.4165-4173.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
- Fujimoto S., Takade A., Amako K., Blaser M. J. Correlation between molecular size of the surface array protein and morphology and antigenicity of the Campylobacter fetus S layer. Infect Immun. 1991 Jun;59(6):2017–2022. doi: 10.1128/iai.59.6.2017-2022.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Guerrant R. L., Lahita R. G., Winn W. C., Jr, Roberts R. B. Campylobacteriosis in man: pathogenic mechanisms and review of 91 bloodstream infections. Am J Med. 1978 Oct;65(4):584–592. doi: 10.1016/0002-9343(78)90845-8. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- McCoy E. C., Doyle D., Burda K., Corbeil L. B., Winter A. J. Superficial antigens of Campylobacter (Vibrio) fetus: characterization of antiphagocytic component. Infect Immun. 1975 Mar;11(3):517–525. doi: 10.1128/iai.11.3.517-525.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pei Z., Blaser M. J. Pathogenesis of Campylobacter fetus infections. Role of surface array proteins in virulence in a mouse model. J Clin Invest. 1990 Apr;85(4):1036–1043. doi: 10.1172/JCI114533. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pei Z., Ellison R. T., 3rd, Lewis R. V., Blaser M. J. Purification and characterization of a family of high molecular weight surface-array proteins from Campylobacter fetus. J Biol Chem. 1988 May 5;263(13):6416–6420. [PubMed] [Google Scholar]
- Plasterk R. H., Simon M. I., Barbour A. G. Transposition of structural genes to an expression sequence on a linear plasmid causes antigenic variation in the bacterium Borrelia hermsii. Nature. 1985 Nov 21;318(6043):257–263. doi: 10.1038/318257a0. [DOI] [PubMed] [Google Scholar]
- Plasterk R. H., Van de Putte P. Genetic switches by DNA inversions in prokaryotes. Biochim Biophys Acta. 1984 Jun 16;782(2):111–119. doi: 10.1016/0167-4781(84)90013-7. [DOI] [PubMed] [Google Scholar]
- Ponticelli A. S., Schultz D. W., Taylor A. F., Smith G. R. Chi-dependent DNA strand cleavage by RecBC enzyme. Cell. 1985 May;41(1):145–151. doi: 10.1016/0092-8674(85)90069-8. [DOI] [PubMed] [Google Scholar]
- 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]
- Seifert H. S., So M. Genetic mechanisms of bacterial antigenic variation. Microbiol Rev. 1988 Sep;52(3):327–336. doi: 10.1128/mr.52.3.327-336.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith G. R. Mechanism and control of homologous recombination in Escherichia coli. Annu Rev Genet. 1987;21:179–201. doi: 10.1146/annurev.ge.21.120187.001143. [DOI] [PubMed] [Google Scholar]
- Stern A., Meyer T. F. Common mechanism controlling phase and antigenic variation in pathogenic neisseriae. Mol Microbiol. 1987 Jul;1(1):5–12. doi: 10.1111/j.1365-2958.1987.tb00520.x. [DOI] [PubMed] [Google Scholar]
- 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]
- Tummuru M. K., Blaser M. J. Characterization of the Campylobacter fetus sapA promoter: evidence that the sapA promoter is deleted in spontaneous mutant strains. J Bacteriol. 1992 Sep;174(18):5916–5922. doi: 10.1128/jb.174.18.5916-5922.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yang L. Y., Pei Z. H., Fujimoto S., Blaser M. J. Reattachment of surface array proteins to Campylobacter fetus cells. J Bacteriol. 1992 Feb;174(4):1258–1267. doi: 10.1128/jb.174.4.1258-1267.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]