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. 1992 Oct;132(2):311–324. doi: 10.1093/genetics/132.2.311

The Relapsing Fever Agent Borrelia Hermsii Has Multiple Copies of Its Chromosome and Linear Plasmids

T Kitten 1, A G Barbour 1
PMCID: PMC1205138  PMID: 1427031

Abstract

Borrelia hermsii, a spirochete which causes relapsing fever in humans and other mammals, eludes the immune response by antigenic variation of the ``Vmp'' proteins. This occurs by replacement of an expressed vmp gene with a copy of a silent vmp gene. Silent and expressed vmp genes are located on separate linear plasmids. To further characterize vmp recombination, copy numbers were determined for two linear plasmids and for the 1-megabase chromosome by comparing hybridization of probes to native DNA with hybridization to recombinant plasmids containing borrelial DNA. Plasmid copy numbers were also estimated by ethidium bromide fluorescence. Total cellular DNA content was determined by spectrophotometry. For borrelias grown in mice, copy numbers and 95% confidence intervals were 14 (12-17) for an expression plasmid, 8 (7-9) for a silent plasmid, and 16 (13-18) for the chromosome. Borrelias grown in broth medium had one-fourth to one-half this number of plasmids and chromosomes. Staining of cells with 4',6-diamidino-2-phenylindole revealed DNA to be distributed throughout most of the spirochete's length. These findings indicate that borrelias organize their total cellular DNA into several complete genomes and that cells undergoing serotype switches do one or more of the following: (1) coexpress Vmps from switched and unswitched expression plasmids for at least three to five generations, (2) suppress transcription from some expression plasmid copies, or (3) partition expression plasmids nonrandomly. The lower copy number of the silent plasmid indicates that nonreciprocal Vmp gene recombination may result from loss of recombinant silent plasmids by segregation.

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Selected References

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  1. Auran N. E., Johnson R. C., Ritzi D. M. Isolation of the outer sheath of Leptospira and its immunogenic properties in hamsters. Infect Immun. 1972 Jun;5(6):968–975. doi: 10.1128/iai.5.6.968-975.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barbour A. G. Antigenic variation of a relapsing fever Borrelia species. Annu Rev Microbiol. 1990;44:155–171. doi: 10.1146/annurev.mi.44.100190.001103. [DOI] [PubMed] [Google Scholar]
  3. Barbour A. G., Burman N., Carter C. J., Kitten T., Bergström S. Variable antigen genes of the relapsing fever agent Borrelia hermsii are activated by promoter addition. Mol Microbiol. 1991 Feb;5(2):489–493. doi: 10.1111/j.1365-2958.1991.tb02132.x. [DOI] [PubMed] [Google Scholar]
  4. Barbour A. G., Carter C. J., Burman N., Freitag C. S., Garon C. F., Bergström S. Tandem insertion sequence-like elements define the expression site for variable antigen genes of Borrelia hermsii. Infect Immun. 1991 Jan;59(1):390–397. doi: 10.1128/iai.59.1.390-397.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Barbour A. G., Garon C. F. Linear plasmids of the bacterium Borrelia burgdorferi have covalently closed ends. Science. 1987 Jul 24;237(4813):409–411. doi: 10.1126/science.3603026. [DOI] [PubMed] [Google Scholar]
  6. Barbour A. G., Hayes S. F. Biology of Borrelia species. Microbiol Rev. 1986 Dec;50(4):381–400. doi: 10.1128/mr.50.4.381-400.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Barbour A. G., Hayes S. F., Heiland R. A., Schrumpf M. E., Tessier S. L. A Borrelia-specific monoclonal antibody binds to a flagellar epitope. Infect Immun. 1986 May;52(2):549–554. doi: 10.1128/iai.52.2.549-554.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Barbour A. G. Isolation and cultivation of Lyme disease spirochetes. Yale J Biol Med. 1984 Jul-Aug;57(4):521–525. [PMC free article] [PubMed] [Google Scholar]
  9. Barbour A. G., Tessier S. L., Stoenner H. G. Variable major proteins of Borrellia hermsii. J Exp Med. 1982 Nov 1;156(5):1312–1324. doi: 10.1084/jem.156.5.1312. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Barbour A. G., Tessier S. L., Todd W. J. Lyme disease spirochetes and ixodid tick spirochetes share a common surface antigenic determinant defined by a monoclonal antibody. Infect Immun. 1983 Aug;41(2):795–804. doi: 10.1128/iai.41.2.795-804.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Barstad P. A., Coligan J. E., Raum M. G., Barbour A. G. Variable major proteins of Borrelia hermsii. Epitope mapping and partial sequence analysis of CNBr peptides. J Exp Med. 1985 Jun 1;161(6):1302–1314. doi: 10.1084/jem.161.6.1302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Borst P. Molecular genetics of antigenic variation. Immunol Today. 1991 Mar;12(3):A29–A33. doi: 10.1016/S0167-5699(05)80009-X. [DOI] [PubMed] [Google Scholar]
  14. Burman N., Bergström S., Restrepo B. I., Barbour A. G. The variable antigens Vmp7 and Vmp21 of the relapsing fever bacterium Borrelia hermsii are structurally analogous to the VSG proteins of the African trypanosome. Mol Microbiol. 1990 Oct;4(10):1715–1726. doi: 10.1111/j.1365-2958.1990.tb00549.x. [DOI] [PubMed] [Google Scholar]
  15. Davidson B. E., MacDougall J., Saint Girons I. Physical map of the linear chromosome of the bacterium Borrelia burgdorferi 212, a causative agent of Lyme disease, and localization of rRNA genes. J Bacteriol. 1992 Jun;174(11):3766–3774. doi: 10.1128/jb.174.11.3766-3774.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Daxhelet G. A., Coene M. M., Hoet P. P., Cocito C. G. Spectrofluorometry of dyes with DNAs of different base composition and conformation. Anal Biochem. 1989 Jun;179(2):401–403. doi: 10.1016/0003-2697(89)90152-8. [DOI] [PubMed] [Google Scholar]
  17. Donachie W. D., Begg K. J. Cell length, nucleoid separation, and cell division of rod-shaped and spherical cells of Escherichia coli. J Bacteriol. 1989 Sep;171(9):4633–4639. doi: 10.1128/jb.171.9.4633-4639.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Engberg B., Nordström K. Replication of R-factor R1 in Scherichia coli K-12 at different growth rates. J Bacteriol. 1975 Jul;123(1):179–186. doi: 10.1128/jb.123.1.179-186.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. FULTON J. D., SMITH P. J. Carbohydrate metabolism in Spirochaeta recurrentis. 1. The metabolism of spirochaetes in vivo and in vitro. Biochem J. 1960 Sep;76:491–499. doi: 10.1042/bj0760491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ferdows M. S., Barbour A. G. Megabase-sized linear DNA in the bacterium Borrelia burgdorferi, the Lyme disease agent. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5969–5973. doi: 10.1073/pnas.86.15.5969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Freeman S. E., Larcom L. L., Thompson B. D. Electrophoretic separation of nucleic acids: evaluation by video and photographic densitometry. Electrophoresis. 1990 May;11(5):425–431. doi: 10.1002/elps.1150110513. [DOI] [PubMed] [Google Scholar]
  22. Fritz R. B., Musich P. R. Unexpected loss of genomic DNA from agarose gel plugs. Biotechniques. 1990 Nov;9(5):542, 544, 546-50. [PubMed] [Google Scholar]
  23. Garon C. F., Dorward D. W., Corwin M. D. Structural features of Borrelia burgdorferi--the Lyme disease spirochete: silver staining for nucleic acids. Scanning Microsc Suppl. 1989;3:109–115. [PubMed] [Google Scholar]
  24. Guerry P., Logan S. M., Thornton S., Trust T. J. Genomic organization and expression of Campylobacter flagellin genes. J Bacteriol. 1990 Apr;172(4):1853–1860. doi: 10.1128/jb.172.4.1853-1860.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  26. Hansen M. T. Multiplicity of genome equivalents in the radiation-resistant bacterium Micrococcus radiodurans. J Bacteriol. 1978 Apr;134(1):71–75. doi: 10.1128/jb.134.1.71-75.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Hedge P. J., Spratt B. G. Amino acid substitutions that reduce the affinity of penicillin-binding protein 3 of Escherichia coli for cephalexin. Eur J Biochem. 1985 Aug 15;151(1):111–121. doi: 10.1111/j.1432-1033.1985.tb09075.x. [DOI] [PubMed] [Google Scholar]
  28. Hiraga S., Niki H., Ogura T., Ichinose C., Mori H., Ezaki B., Jaffé A. Chromosome partitioning in Escherichia coli: novel mutants producing anucleate cells. J Bacteriol. 1989 Mar;171(3):1496–1505. doi: 10.1128/jb.171.3.1496-1505.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Holliday R. Genomic imprinting and allelic exclusion. Dev Suppl. 1990:125–129. [PubMed] [Google Scholar]
  30. Hyde F. W., Johnson R. C. Genetic relationship of lyme disease spirochetes to Borrelia, Treponema, and Leptospira spp. J Clin Microbiol. 1984 Aug;20(2):151–154. doi: 10.1128/jcm.20.2.151-154.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Keasling J. D., Palsson B. O., Cooper S. Cell-cycle-specific F plasmid replication: regulation by cell size control of initiation. J Bacteriol. 1991 Apr;173(8):2673–2680. doi: 10.1128/jb.173.8.2673-2680.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Kitten T., Barbour A. G. Juxtaposition of expressed variable antigen genes with a conserved telomere in the bacterium Borrelia hermsii. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6077–6081. doi: 10.1073/pnas.87.16.6077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Kontomichalou P., Mitani M., Clowes R. C. Circular R-factor molecules controlling penicillinase synthesis, replicating in Escherichia coli under either relaxed or stringent control. J Bacteriol. 1970 Oct;104(1):34–44. doi: 10.1128/jb.104.1.34-44.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. LePecq J. B., Paoletti C. A fluorescent complex between ethidium bromide and nucleic acids. Physical-chemical characterization. J Mol Biol. 1967 Jul 14;27(1):87–106. doi: 10.1016/0022-2836(67)90353-1. [DOI] [PubMed] [Google Scholar]
  35. Meier J. T., Simon M. I., Barbour A. G. Antigenic variation is associated with DNA rearrangements in a relapsing fever Borrelia. Cell. 1985 Jun;41(2):403–409. doi: 10.1016/s0092-8674(85)80013-1. [DOI] [PubMed] [Google Scholar]
  36. Meinkoth J., Wahl G. Hybridization of nucleic acids immobilized on solid supports. Anal Biochem. 1984 May 1;138(2):267–284. doi: 10.1016/0003-2697(84)90808-x. [DOI] [PubMed] [Google Scholar]
  37. Nagpal P., Jafri S., Reddy M. A., Das H. K. Multiple chromosomes of Azotobacter vinelandii. J Bacteriol. 1989 Jun;171(6):3133–3138. doi: 10.1128/jb.171.6.3133-3138.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. O'Neill R. R., Mitchell L. G., Merril C. R., Rasband W. S. Use of image analysis to quantitate changes in form of mitochondrial DNA after x-irradiation. Appl Theor Electrophor. 1989;1(3):163–167. [PubMed] [Google Scholar]
  39. Paster B. J., Dewhirst F. E., Weisburg W. G., Tordoff L. A., Fraser G. J., Hespell R. B., Stanton T. B., Zablen L., Mandelco L., Woese C. R. Phylogenetic analysis of the spirochetes. J Bacteriol. 1991 Oct;173(19):6101–6109. doi: 10.1128/jb.173.19.6101-6109.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. 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]
  41. Pleier E., Schmitt R. Identification and sequence analysis of two related flagellin genes in Rhizobium meliloti. J Bacteriol. 1989 Mar;171(3):1467–1475. doi: 10.1128/jb.171.3.1467-1475.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Prentki P., Chandler M., Caro L. Replication of prophage P1 during the cell cycle of Escherichia coli. Mol Gen Genet. 1977 Mar 28;152(1):71–76. doi: 10.1007/BF00264942. [DOI] [PubMed] [Google Scholar]
  43. Projan S. J., Carleton S., Novick R. P. Determination of plasmid copy number by fluorescence densitometry. Plasmid. 1983 Mar;9(2):182–190. doi: 10.1016/0147-619x(83)90019-7. [DOI] [PubMed] [Google Scholar]
  44. Ribeiro E. A., Larcom L. L., Miller D. P. Quantitative fluorescence of DNA-intercalated ethidium bromide on agarose gels. Anal Biochem. 1989 Sep;181(2):197–208. doi: 10.1016/0003-2697(89)90229-7. [DOI] [PubMed] [Google Scholar]
  45. Ricard M., Hirota Y. Process of cellular division in Escherichia coli: physiological study on thermosensitive mutants defective in cell division. J Bacteriol. 1973 Oct;116(1):314–322. doi: 10.1128/jb.116.1.314-322.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Rownd R. Replication of a bacterial episome under relaxed control. J Mol Biol. 1969 Sep 28;44(3):387–402. doi: 10.1016/0022-2836(69)90368-4. [DOI] [PubMed] [Google Scholar]
  47. Sadoff H. L., Shimel B., Ellis S. Characterization of Azotobacter vinelandii deoxyribonucleic acid and folded chromosomes. J Bacteriol. 1979 Jun;138(3):871–877. doi: 10.1128/jb.138.3.871-877.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Stoenner H. G., Dodd T., Larsen C. Antigenic variation of Borrelia hermsii. J Exp Med. 1982 Nov 1;156(5):1297–1311. doi: 10.1084/jem.156.5.1297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. 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]
  50. Van der Ploeg L. H., Smith C. L., Polvere R. I., Gottesdiener K. M. Improved separation of chromosome-sized DNA from Trypanosoma brucei, stock 427-60. Nucleic Acids Res. 1989 Apr 25;17(8):3217–3227. doi: 10.1093/nar/17.8.3217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]

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