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. 1997 Dec;179(23):7523–7529. doi: 10.1128/jb.179.23.7523-7529.1997

Nested DNA inversion of Campylobacter fetus S-layer genes is recA dependent.

J Dworkin 1, O L Shedd 1, M J Blaser 1
PMCID: PMC179705  PMID: 9393719

Abstract

Wild-type strains of Campylobacter fetus are covered by a monomolecular array of surface layer proteins (SLPs) critical for virulence. Each cell possesses eight SLP gene cassettes, tightly clustered in the genome, that encode SLPs of 97 to 149 kDa. Variation of SLP expression occurs by a mechanism of nested DNA rearrangement that involves the inversion of a 6.2-kb sapA promoter-containing element alone or together with one or more flanking SLP gene cassettes. The presence of extensive regions of identity flanking the 5' and 3' ends of each SLP gene cassette and of a Chi-like recognition sequence within the 5' region of identity suggests that rearrangement of SLP gene cassettes may occur by a generalized (RecA-dependent) homologous recombination pathway. To explore this possibility, we cloned C. fetus recA and created mutant strains by marker rescue, in which recA is disrupted in either S+ or S- strains. These mutants then were assessed for their abilities to alter SLP expression either in the presence or absence of a complementary shuttle plasmid harboring native recA. In contrast to all previously reported programmed DNA inversion systems, inversion in C. fetus is recA dependent.

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Selected References

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  1. 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]
  2. 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]
  3. 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]
  4. 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]
  5. 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]
  6. Blaser M. J., Wang E., Tummuru M. K., Washburn R., Fujimoto S., Labigne A. High-frequency S-layer protein variation in Campylobacter fetus revealed by sapA mutagenesis. Mol Microbiol. 1994 Nov;14(3):453–462. doi: 10.1111/j.1365-2958.1994.tb02180.x. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Craig N. L. The mechanism of conservative site-specific recombination. Annu Rev Genet. 1988;22:77–105. doi: 10.1146/annurev.ge.22.120188.000453. [DOI] [PubMed] [Google Scholar]
  9. Dworkin J., Blaser M. J. Generation of Campylobacter fetus S-layer protein diversity utilizes a single promoter on an invertible DNA segment. Mol Microbiol. 1996 Mar;19(6):1241–1253. doi: 10.1111/j.1365-2958.1996.tb02469.x. [DOI] [PubMed] [Google Scholar]
  10. Dworkin J., Blaser M. J. Nested DNA inversion as a paradigm of programmed gene rearrangement. Proc Natl Acad Sci U S A. 1997 Feb 4;94(3):985–990. doi: 10.1073/pnas.94.3.985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dworkin J., Tummuru M. K., Blaser M. J. A lipopolysaccharide-binding domain of the Campylobacter fetus S-layer protein resides within the conserved N terminus of a family of silent and divergent homologs. J Bacteriol. 1995 Apr;177(7):1734–1741. doi: 10.1128/jb.177.7.1734-1741.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dworkin J., Tummuru M. K., Blaser M. J. Segmental conservation of sapA sequences in type B Campylobacter fetus cells. J Biol Chem. 1995 Jun 23;270(25):15093–15101. doi: 10.1074/jbc.270.25.15093. [DOI] [PubMed] [Google Scholar]
  13. Dybvig K. DNA rearrangements and phenotypic switching in prokaryotes. Mol Microbiol. 1993 Nov;10(3):465–471. doi: 10.1111/j.1365-2958.1993.tb00919.x. [DOI] [PubMed] [Google Scholar]
  14. Ennis D. G., Amundsen S. K., Smith G. R. Genetic functions promoting homologous recombination in Escherichia coli: a study of inversions in phage lambda. Genetics. 1987 Jan;115(1):11–24. doi: 10.1093/genetics/115.1.11. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Figurski D., Meyer R., Miller D. S., Helinski D. R. Generation in vitro of deletions in the broad host range plasmid RK2 using phage Mu insertions and a restriction endonuclease. Gene. 1976;1(1):107–119. doi: 10.1016/0378-1119(76)90010-x. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Garcia M. M., Lutze-Wallace C. L., Denes A. S., Eaglesome M. D., Holst E., Blaser M. J. Protein shift and antigenic variation in the S-layer of Campylobacter fetus subsp. venerealis during bovine infection accompanied by genomic rearrangement of sapA homologs. J Bacteriol. 1995 Apr;177(8):1976–1980. doi: 10.1128/jb.177.8.1976-1980.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Guerry P., Pope P. M., Burr D. H., Leifer J., Joseph S. W., Bourgeois A. L. Development and characterization of recA mutants of Campylobacter jejuni for inclusion in attenuated vaccines. Infect Immun. 1994 Feb;62(2):426–432. doi: 10.1128/iai.62.2.426-432.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hagemann A. T., Craig N. L. Tn7 transposition creates a hotspot for homologous recombination at the transposon donor site. Genetics. 1993 Jan;133(1):9–16. doi: 10.1093/genetics/133.1.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hiestand-Nauer R., Iida S. Sequence of the site-specific recombinase gene cin and of its substrates serving in the inversion of the C segment of bacteriophage P1. EMBO J. 1983;2(10):1733–1740. doi: 10.1002/j.1460-2075.1983.tb01650.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hill C. W., Harnish B. W. Inversions between ribosomal RNA genes of Escherichia coli. Proc Natl Acad Sci U S A. 1981 Nov;78(11):7069–7072. doi: 10.1073/pnas.78.11.7069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Johnson R. C., Bruist M. F., Simon M. I. Host protein requirements for in vitro site-specific DNA inversion. Cell. 1986 Aug 15;46(4):531–539. doi: 10.1016/0092-8674(86)90878-0. [DOI] [PubMed] [Google Scholar]
  23. Kanaar R., van de Putte P. Topological aspects of site-specific DNA-inversion. Bioessays. 1987 Nov;7(5):195–200. doi: 10.1002/bies.950070502. [DOI] [PubMed] [Google Scholar]
  24. Kostriken R., Strathern J. N., Klar A. J., Hicks J. B., Heffron F. A site-specific endonuclease essential for mating-type switching in Saccharomyces cerevisiae. Cell. 1983 Nov;35(1):167–174. doi: 10.1016/0092-8674(83)90219-2. [DOI] [PubMed] [Google Scholar]
  25. Kuzminov A., Schabtach E., Stahl F. W. Chi sites in combination with RecA protein increase the survival of linear DNA in Escherichia coli by inactivating exoV activity of RecBCD nuclease. EMBO J. 1994 Jun 15;13(12):2764–2776. doi: 10.1002/j.1460-2075.1994.tb06570.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Labigne-Roussel A., Courcoux P., Tompkins L. Gene disruption and replacement as a feasible approach for mutagenesis of Campylobacter jejuni. J Bacteriol. 1988 Apr;170(4):1704–1708. doi: 10.1128/jb.170.4.1704-1708.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Labigne-Roussel A., Harel J., Tompkins L. Gene transfer from Escherichia coli to Campylobacter species: development of shuttle vectors for genetic analysis of Campylobacter jejuni. J Bacteriol. 1987 Nov;169(11):5320–5323. doi: 10.1128/jb.169.11.5320-5323.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Labigne A., Cussac V., Courcoux P. Shuttle cloning and nucleotide sequences of Helicobacter pylori genes responsible for urease activity. J Bacteriol. 1991 Mar;173(6):1920–1931. doi: 10.1128/jb.173.6.1920-1931.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Manavathu E. K., Hiratsuka K., Taylor D. E. Nucleotide sequence analysis and expression of a tetracycline-resistance gene from Campylobacter jejuni. Gene. 1988;62(1):17–26. doi: 10.1016/0378-1119(88)90576-8. [DOI] [PubMed] [Google Scholar]
  30. Mertens G., Hoffmann A., Blöcker H., Frank R., Kahmann R. Gin-mediated site-specific recombination in bacteriophage Mu DNA: overproduction of the protein and inversion in vitro. EMBO J. 1984 Oct;3(10):2415–2421. doi: 10.1002/j.1460-2075.1984.tb02148.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Murphy E., Novick R. P. Physical mapping of Staphylococcus aureus penicillinase plasmid pI524: characterization of an invertible region. Mol Gen Genet. 1979 Aug;175(1):19–30. doi: 10.1007/BF00267851. [DOI] [PubMed] [Google Scholar]
  32. Murphy E., Novick R. P. Site-specific recombination between plasmids of Staphylococcus aureus. J Bacteriol. 1980 Jan;141(1):316–326. doi: 10.1128/jb.141.1.316-326.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Paulsen I. T., Gillespie M. T., Littlejohn T. G., Hanvivatvong O., Rowland S. J., Dyke K. G., Skurray R. A. Characterisation of sin, a potential recombinase-encoding gene from Staphylococcus aureus. Gene. 1994 Apr 8;141(1):109–114. doi: 10.1016/0378-1119(94)90136-8. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. 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]
  36. Petes T. D., Hill C. W. Recombination between repeated genes in microorganisms. Annu Rev Genet. 1988;22:147–168. doi: 10.1146/annurev.ge.22.120188.001051. [DOI] [PubMed] [Google Scholar]
  37. 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]
  38. Roca A. I., Cox M. M. The RecA protein: structure and function. Crit Rev Biochem Mol Biol. 1990;25(6):415–456. doi: 10.3109/10409239009090617. [DOI] [PubMed] [Google Scholar]
  39. Sadowski P. Site-specific recombinases: changing partners and doing the twist. J Bacteriol. 1986 Feb;165(2):341–347. doi: 10.1128/jb.165.2.341-347.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Saldanha R., Mohr G., Belfort M., Lambowitz A. M. Group I and group II introns. FASEB J. 1993 Jan;7(1):15–24. doi: 10.1096/fasebj.7.1.8422962. [DOI] [PubMed] [Google Scholar]
  41. Sandmeier H. Acquisition and rearrangement of sequence motifs in the evolution of bacteriophage tail fibres. Mol Microbiol. 1994 May;12(3):343–350. doi: 10.1111/j.1365-2958.1994.tb01023.x. [DOI] [PubMed] [Google Scholar]
  42. Schmid M. B., Roth J. R. Selection and endpoint distribution of bacterial inversion mutations. Genetics. 1983 Nov;105(3):539–557. doi: 10.1093/genetics/105.3.539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Schofield M. A., Agbunag R., Miller J. H. DNA inversions between short inverted repeats in Escherichia coli. Genetics. 1992 Oct;132(2):295–302. doi: 10.1093/genetics/132.2.295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Silverman M., Zieg J., Hilmen M., Simon M. Phase variation in Salmonella: genetic analysis of a recombinational switch. Proc Natl Acad Sci U S A. 1979 Jan;76(1):391–395. doi: 10.1073/pnas.76.1.391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Smith G. R. Homologous recombination in procaryotes. Microbiol Rev. 1988 Mar;52(1):1–28. doi: 10.1128/mr.52.1.1-28.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Strathern J. N., Klar A. J., Hicks J. B., Abraham J. A., Ivy J. M., Nasmyth K. A., McGill C. Homothallic switching of yeast mating type cassettes is initiated by a double-stranded cut in the MAT locus. Cell. 1982 Nov;31(1):183–192. doi: 10.1016/0092-8674(82)90418-4. [DOI] [PubMed] [Google Scholar]
  47. Szostak J. W., Orr-Weaver T. L., Rothstein R. J., Stahl F. W. The double-strand-break repair model for recombination. Cell. 1983 May;33(1):25–35. doi: 10.1016/0092-8674(83)90331-8. [DOI] [PubMed] [Google Scholar]
  48. Thaler D. S., Stahl F. W. DNA double-chain breaks in recombination of phage lambda and of yeast. Annu Rev Genet. 1988;22:169–197. doi: 10.1146/annurev.ge.22.120188.001125. [DOI] [PubMed] [Google Scholar]
  49. Thompson S. A., Blaser M. J. Isolation of the Helicobacter pylori recA gene and involvement of the recA region in resistance to low pH. Infect Immun. 1995 Jun;63(6):2185–2193. doi: 10.1128/iai.63.6.2185-2193.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. 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]
  51. Tummuru M. K., Blaser M. J. Rearrangement of sapA homologs with conserved and variable regions in Campylobacter fetus. Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):7265–7269. doi: 10.1073/pnas.90.15.7265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Wang E., Garcia M. M., Blake M. S., Pei Z., Blaser M. J. Shift in S-layer protein expression responsible for antigenic variation in Campylobacter fetus. J Bacteriol. 1993 Aug;175(16):4979–4984. doi: 10.1128/jb.175.16.4979-4984.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Wang Y., Taylor D. E. Chloramphenicol resistance in Campylobacter coli: nucleotide sequence, expression, and cloning vector construction. Gene. 1990 Sep 28;94(1):23–28. doi: 10.1016/0378-1119(90)90463-2. [DOI] [PubMed] [Google Scholar]
  54. White C. I., Haber J. E. Intermediates of recombination during mating type switching in Saccharomyces cerevisiae. EMBO J. 1990 Mar;9(3):663–673. doi: 10.1002/j.1460-2075.1990.tb08158.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Winter A. J., McCoy E. C., Fullmer C. S., Burda K., Bier P. J. Microcapsule of Campylobacter fetus: chemical and physical characterization. Infect Immun. 1978 Dec;22(3):963–971. doi: 10.1128/iai.22.3.963-971.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. 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]
  57. van de Putte P., Goosen N. DNA inversions in phages and bacteria. Trends Genet. 1992 Dec;8(12):457–462. doi: 10.1016/0168-9525(92)90331-w. [DOI] [PubMed] [Google Scholar]

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