The Relapsing Fever Spirochete Borrelia hermsiiContains Multiple, Antigen-Encoding Circular Plasmids That Are Homologous to the cp32 Plasmids of Lyme Disease Spirochetes

ABSTRACT Borrelia hermsii, an agent of tick-borne relapsing fever, was found to contain multiple circular plasmids approximately 30 kb in size. Sequencing of a DNA library constructed from circular plasmid fragments enabled assembly of a composite DNA sequence that is homologous to the cp32 plasmid family of the Lyme disease spirochete,B. burgdorferi. Analysis of another relapsing fever bacterium, B. parkeri, indicated that it contains linear homologs of the B. hermsii and B. burgdorfericp32 plasmids. The B. hermsii cp32 plasmids encode homologs of the B. burgdorferi Mlp and Bdr antigenic proteins and BlyA/BlyB putative hemolysins, but homologs of B. burgdorferi erp genes were absent. Immunoblot analyses demonstrated that relapsing fever patients produced antibodies to Mlp proteins, indicating that those proteins are synthesized by the spirochetes during human infection. Conservation of cp32-encoded genes in differentBorrelia species suggests that their protein products serve functions essential to both relapsing fever and Lyme disease spirochetes. Relapsing fever borreliae replicate to high levels in the blood of infected animals, permitting direct detection and possible functional studies of Mlp, Bdr, BlyA/BlyB, and other cp32-encoded proteins in vivo.

[1]  T. Schwan,et al.  Borrelia burgdorferi contains repeated DNA sequences that are species specific and plasmid associated , 1990, Infection and immunity.

[2]  J. Radolf,et al.  Molecular and Evolutionary Analysis ofBorrelia burgdorferi 297 Circular Plasmid-Encoded Lipoproteins with OspE- and OspF-Like Leader Peptides , 1999, Infection and Immunity.

[3]  B. Stevenson,et al.  Stability of erp Loci duringBorrelia burgdorferi Infection: Recombination Is Not Required for Chronic Infection of Immunocompetent Mice , 1999, Infection and Immunity.

[4]  T. Schwan,et al.  Identification of a protein in several Borrelia species which is related to OspC of the Lyme disease spirochetes , 1993, Journal of clinical microbiology.

[5]  A. Barbour,et al.  The nucleotide sequence of a linear plasmid of Borrelia burgdorferi reveals similarities to those of circular plasmids of other prokaryotes , 1996, Journal of bacteriology.

[6]  T. Schwan,et al.  Changes in infectivity and plasmid profile of the Lyme disease spirochete, Borrelia burgdorferi, as a result of in vitro cultivation , 1988, Infection and immunity.

[7]  Albert Balows,et al.  Manual of Clinical Microbiology, 7th ed. , 2000 .

[8]  S. Casjens,et al.  Evidence of past recombination events among the genes encoding the Erp antigens of Borrelia burgdorferi. , 1998, Microbiology.

[9]  T. Schwan,et al.  Temperature-related differential expression of antigens in the Lyme disease spirochete, Borrelia burgdorferi , 1995, Infection and immunity.

[10]  O. White,et al.  A bacterial genome in flux: the twelve linear and nine circular extrachromosomal DNAs in an infectious isolate of the Lyme disease spirochete Borrelia burgdorferi , 2000, Molecular microbiology.

[11]  T. Schwan,et al.  Temporal Changes in Outer Surface Proteins A and C of the Lyme Disease-Associated Spirochete, Borrelia burgdorferi, during the Chain of Infection in Ticks and Mice , 2000, Journal of Clinical Microbiology.

[12]  W. Zückert,et al.  Circular and linear plasmids of Lyme disease spirochetes have extensive homology: characterization of a repeated DNA element , 1996, Journal of bacteriology.

[13]  W. Burgdorfer,et al.  Outbreak of tick-borne relapsing fever in Spokane County, Washington. , 1969, JAMA.

[14]  B. Stevenson,et al.  Borrelia burgdorferi B31 Erp Proteins That Are Dominant Immunoblot Antigens of Animals Infected with Isolate B31 Are Recognized by Only a Subset of Human Lyme Disease Patient Sera , 2000, Journal of Clinical Microbiology.

[15]  S. Norris,et al.  Antigenic Variation in Lyme Disease Borreliae by Promiscuous Recombination of VMP-like Sequence Cassettes , 1997, Cell.

[16]  B. Luft,et al.  Complete nucleotide sequence of a circular plasmid from the Lyme disease spirochete, Borrelia burgdorferi , 1994, Journal of bacteriology.

[17]  T. Schwan,et al.  Population Structure of the Relapsing Fever Spirochete Borrelia hermsii as Indicated by Polymorphism of Two Multigene Families That Encode Immunogenic Outer Surface Lipoproteins , 1998, Infection and Immunity.

[18]  R. C. Johnson,et al.  Analysis and comparison of plasmid profiles of Borrelia burgdorferi sensu lato strains , 1995, Journal of clinical microbiology.

[19]  D. E. Anderson,et al.  Tick-borne relapsing fever in the northwestern United States and southwestern Canada. , 1998, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[20]  E. Fikrig,et al.  Borrelia burgdorferi OspA is an arthropod-specific transmission- blocking Lyme disease vaccine , 1996, The Journal of experimental medicine.

[21]  A. Hofmann,et al.  Immunological and molecular polymorphisms of OspC, an immunodominant major outer surface protein of Borrelia burgdorferi , 1993, Infection and immunity.

[22]  C. Garon,et al.  Coumermycin A1 inhibits growth and induces relaxation of supercoiled plasmids in Borrelia burgdorferi, the Lyme disease agent , 1993, Antimicrobial Agents and Chemotherapy.

[23]  M. Simon,et al.  Antigenic variation is associated with DNA rearrangements in a relapsing fever borrelia , 1985, Cell.

[24]  J. W. Chase,et al.  Single‐stranded DNA binding proteins (SSBs) from prokaryotic transmissible plasmids , 1991, Proteins.

[25]  J. Radolf,et al.  Identification, Characterization, and Expression of Three New Members of the Borrelia burgdorferi Mlp (2.9) Lipoprotein Gene Family , 1999, Infection and Immunity.

[26]  E. Fikrig,et al.  Borrelia burgdorferi genes selectively expressed in the infected host. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[27]  A. Barbour,et al.  Plasmid analysis of Borrelia burgdorferi, the Lyme disease agent , 1988, Journal of clinical microbiology.

[28]  T. Schwan.  Ticks and Borrelia: model systems for investigating pathogen-arthropod interactions. , 1996, Infectious agents and disease.

[29]  J. Bono,et al.  Characterization of cp18, a naturally truncated member of the cp32 family of Borrelia burgdorferi plasmids , 1997, Journal of bacteriology.

[30]  A. Barbour,et al.  Antigenic variation of a relapsing fever Borrelia species. , 1990, Annual review of microbiology.

[31]  M. Kramer,et al.  Molecular cloning and immunological characterization of a novel linear-plasmid-encoded gene, pG, of Borrelia burgdorferi expressed only in vivo , 1995, Infection and immunity.

[32]  A. Barbour.  Isolation and cultivation of Lyme disease spirochetes. , 1984, The Yale journal of biology and medicine.

[33]  Functional Activities of Antibodies Directed against Surface Lipoproteins of Borrelia hermsii , 1995, Microbiology and immunology.

[34]  L. Gern,et al.  Expression of outer surface proteins A and C of Borrelia afzelii in Ixodes ricinus ticks and in the skin of mice. , 1998, Zentralblatt fur Bakteriologie : international journal of medical microbiology.

[35]  J. Radolf,et al.  Evidence for in vivo but not in vitro expression of a Borrelia burgdorferi outer surface protein F (OspF) homologue , 1995, Molecular microbiology.

[36]  G. Baranton,et al.  Polymerase chain reaction with the 30-kb circular plasmid of Borrelia burgdorferi B31 as a target for detection of the Lyme borreliosis agents in cerebrospinal fluid. , 1993, Research in microbiology.

[37]  J. Hurwitz,et al.  Methylation of DNA , 1966, The Journal of general physiology.

[38]  J. D. Bendtsen,et al.  Identification and Characterization of the Single-Stranded DNA-Binding Protein of Bacteriophage P1 , 1999, Journal of bacteriology.

[39]  D. Roberts,et al.  Evolutionary and molecular analyses of the Borrelia bdr super gene family: delineation of distinct sub-families and demonstration of the genus wide conservation of putative functional domains, structural properties and repeat motifs. , 2000, Microbial pathogenesis.

[40]  J. P. Jost,et al.  DNA Methylation: Molecular Biology and Biological Significance , 1993 .

[41]  B. Kagan,et al.  Isolation and characterization of the outer membrane of Borrelia hermsii. , 1998, Infection and immunity.

[42]  Schwan Tg.  Ticks and Borrelia: model systems for investigating pathogen-arthropod interactions. , 1996 .

[43]  M. Li,et al.  Borrelia burgdorferi supercoiled plasmids encode multicopy tandem open reading frames and a lipoprotein gene family , 1996, Journal of bacteriology.

[44]  T. Schwan,et al.  Homology between Borrelia burgdorferi OspC and members of the family of Borrelia hermsii variable major proteins. , 1994, Gene.

[45]  T. Bugge,et al.  Plasminogen Is Required for Efficient Dissemination of B. burgdorferi in Ticks and for Enhancement of Spirochetemia in Mice , 1997, Cell.

[46]  S. Casjens,et al.  Homology throughout the multiple 32-kilobase circular plasmids present in Lyme disease spirochetes , 1997, Journal of bacteriology.

[47]  T. Guina,et al.  Cloning and analysis of a Borrelia burgdorferi membrane‐interactive protein exhibiting haemolytic activity , 1997, Molecular microbiology.

[48]  Christian H. Eggers,et al.  Molecular Evidence for a New Bacteriophage ofBorrelia burgdorferi , 1999, Journal of bacteriology.

[49]  T. Schwan,et al.  Bloodstream- versus tick-associated variants of a relapsing fever bacterium. , 1998, Science.

[50]  D. Sonenshine.  Biology of ticks , 1991 .

[51]  R. C. Johnson,et al.  Methylated DNA in Borrelia species , 1990, Journal of bacteriology.

[52]  J. Radolf,et al.  Molecular and Evolutionary Characterization of the cp32/18 Family of Supercoiled Plasmids in Borrelia burgdorferi297 , 2000, Infection and Immunity.

[53]  B. Stevenson,et al.  A family of genes located on four separate 32-kilobase circular plasmids in Borrelia burgdorferi B31 , 1996, Journal of bacteriology.

[54]  S. Salzberg,et al.  Genomic sequence of a Lyme disease spirochaete, Borrelia burgdorferi , 1997, Nature.

[55]  T. Trautner,et al.  Methylation of DNA in prokaryotes. , 1993, EXS.

[56]  J. Piesman,et al.  Inhibition of Borrelia burgdorferi Migration from the Midgut to the Salivary Glands following Feeding by Ticks on OspC-Immunized Mice , 2000, Infection and Immunity.

[57]  T. Schwan,et al.  Tick-Borne Relapsing Fever in British Columbia, Canada: First Isolation of Borrelia hermsii , 1998, Journal of Clinical Microbiology.

[58]  T. Schwan,et al.  Induction of an outer surface protein on Borrelia burgdorferi during tick feeding. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[59]  S. Casjens,et al.  Linear chromosomal physical and genetic map of Borrelia burgdorferi, the Lyme disease agent , 1993, Molecular microbiology.

[60]  C. Benoist,et al.  Cell Death Mediators in Autoimmune Diabetes—No Shortage of Suspects , 1997, Cell.

[61]  A. Barbour,et al.  Comparative Analysis and Immunological Characterization of the Borrelia Bdr Protein Family , 1999, Infection and Immunity.

[62]  H. Stoenner,et al.  Antigenic variation of Borrelia hermsii , 1982, The Journal of experimental medicine.

[63]  A. Barbour,et al.  Biology of Borrelia species. , 1986, Microbiological reviews.

[64]  J. Bono,et al.  Borrelia burgdorferi Erp Proteins Are Immunogenic in Mammals Infected by Tick Bite, and Their Synthesis Is Inducible in Cultured Bacteria , 1998, Infection and Immunity.

[65]  Ruth R. Montgomery,et al.  Outer surface proteins E and F of Borrelia burgdorferi, the agent of Lyme disease , 1994, Infection and immunity.

[66]  U. Munderloh,et al.  Expression of outer surface proteins A and C of Borrelia burgdorferi in Ixodes ricinus ticks removed from humans , 1998, Medical Microbiology and Immunology.

[67]  J. Carlyon,et al.  Cloning and Molecular Characterization of a Multicopy, Linear Plasmid-Carried, Repeat Motif-Containing Gene from Borrelia turicatae, a Causative Agent of Relapsing Fever , 1998, Journal of bacteriology.