A plasmid‐encoded nicotinamidase (PncA) is essential for infectivity of Borrelia burgdorferi in a mammalian host

Borrelia burgdorferi, a spirochaete that causes Lyme borreliosis, contains 21 linear and circular plasmids thought to be important for survival in mammals or ticks. Our results demonstrate that the gene BBE22 encoding a nicotinamidase is capable of replacing the requirement for the 25 kb linear plasmid lp25 during mammalian infection. Transformation of B. burgdorferi lacking lp25 with a shuttle vector containing the lp25 gene BBE22 (pBBE22) restored infectivity in mice to a level comparable to that of wild‐type Borrelia. This complementation also restored the growth and host adaptation of lp25–B. burgdorferi in dialysis membrane chambers (DMCs) implanted in rats. A single Cys to Ala conversion at the putative active site of BBE22 abrogated the ability of pBBE22 to re‐establish infectivity or growth in DMCs. Additional Salmonella typhimurium complementation studies and enzymatic analysis demonstrated that the BBE22 gene product has nicotinamidase activity and is most probably required for the biosynthesis of NAD. These results indicate that some plasmid‐encoded products fulfil physiological functions required in the enzootic cycle of pathogenic Borrelia.

[1]  S. Norris,et al.  Decreased Electroporation Efficiency in Borrelia burgdorferi Containing Linear Plasmids lp25 and lp56: Impact on Transformation of Infectious B. burgdorferi , 2002, Infection and Immunity.

[2]  S. Wikel,et al.  Changes in Temporal and Spatial Patterns of Outer Surface Lipoprotein Expression Generate Population Heterogeneity and Antigenic Diversity in the Lyme Disease Spirochete, Borrelia burgdorferi , 2002, Infection and Immunity.

[3]  T. Klabunde,et al.  Crystal Structure of Lyme Disease Variable Surface Antigen VlsE of Borrelia burgdorferi * , 2002, The Journal of Biological Chemistry.

[4]  J. Bono,et al.  Clonal Polymorphism of Borrelia burgdorferi Strain B31 MI: Implications for Mutagenesis in an Infectious Strain Background , 2002, Infection and Immunity.

[5]  Andrew T. Revel,et al.  DNA microarray analysis of differential gene expression in Borrelia burgdorferi, the Lyme disease spirochete , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[6]  B. Stevenson,et al.  Differential Binding of Host Complement Inhibitor Factor H by Borrelia burgdorferi Erp Surface Proteins: a Possible Mechanism Underlying the Expansive Host Range of Lyme Disease Spirochetes , 2002, Infection and Immunity.

[7]  J. Radolf,et al.  Identification of loci critical for replication and compatibility of a Borrelia burgdorferi cp32 plasmid and use of a cp32‐based shuttle vector for the expression of fluorescent reporters in the Lyme disease spirochaete , 2002, Molecular microbiology.

[8]  Man-Huei Chang,et al.  Summary of notifiable diseases--United States, 2000. , 2002, MMWR. Morbidity and mortality weekly report.

[9]  S H Kim,et al.  Crystal structure and mechanism of catalysis of a pyrazinamidase from Pyrococcus horikoshii. , 2001, Biochemistry.

[10]  S. Norris,et al.  Analysis of Borrelia burgdorferivlsE Gene Expression and Recombination in the Tick Vector , 2001, Infection and Immunity.

[11]  F. Cabello,et al.  Expression of Borrelia burgdorferi OspC and DbpA is controlled by a RpoN–RpoS regulatory pathway , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[12]  J. Bono,et al.  Genetics and Regulation of Chitobiose Utilization inBorrelia burgdorferi , 2001, Journal of bacteriology.

[13]  J. A. Carroll,et al.  Borrelia burgdorferi RevA Antigen Is a Surface-Exposed Outer Membrane Protein Whose Expression Is Regulated in Response to Environmental Temperature and pH , 2001, Infection and Immunity.

[14]  M. Mbow,et al.  Analysis of Borrelia burgdorferi gene expression during life cycle phases of the tick vector Ixodes scapularis. , 2001, Microbes and infection.

[15]  J. Radolf,et al.  Regulation of OspE-Related, OspF-Related, and Elp Lipoproteins of Borrelia burgdorferi Strain 297 by Mammalian Host-Specific Signals , 2001, Infection and Immunity.

[16]  Pekka Lahdenne,et al.  The Complement Regulator Factor H Binds to the Surface Protein OspE of Borrelia burgdorferi * , 2001, The Journal of Biological Chemistry.

[17]  J. Bono,et al.  Isolation of a circular plasmid region sufficient for autonomous replication and transformation of infectious Borrelia burgdorferi , 2001, Molecular microbiology.

[18]  Maria Labandeira-Rey,et al.  Decreased Infectivity in Borrelia burgdorferi Strain B31 Is Associated with Loss of Linear Plasmid 25 or 28-1 , 2001, Infection and Immunity.

[19]  M. Saier,et al.  Transcriptional regulation in spirochetes of medical importance. , 2001 .

[20]  S. Norris,et al.  Correlation between plasmid content and infectivity in Borrelia burgdorferi. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[21]  J. Skare,et al.  The many faces of Borrelia burgdorferi. , 2000, Journal of molecular microbiology and biotechnology.

[22]  S. Wikel,et al.  Interdependence of environmental factors influencing reciprocal patterns of gene expression in virulent Borrelia burgdorferi , 2000, Molecular microbiology.

[23]  S. Casjens,et al.  Distribution of Twelve Linear Extrachromosomal DNAs in Natural Isolates of Lyme Disease Spirochetes , 2000, Journal of bacteriology.

[24]  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.

[25]  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.

[26]  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.

[27]  J. A. Carroll,et al.  Effects of Environmental pH on Membrane Proteins in Borrelia burgdorferi , 1999, Infection and Immunity.

[28]  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.

[29]  R. Doerge,et al.  Identification of quantitative trait loci governing arthritis severity and humoral responses in the murine model of Lyme disease. , 1999, Journal of immunology.

[30]  M. Hanson,et al.  DbpA, but Not OspA, Is Expressed by Borrelia burgdorferi during Spirochetemia and Is a Target for Protective Antibodies , 1998, Infection and Immunity.

[31]  C Colovos,et al.  The 1.8 A crystal structure of the ycaC gene product from Escherichia coli reveals an octameric hydrolase of unknown specificity. , 1998, Structure.

[32]  S. Norris,et al.  Genetic Variation of the Borrelia burgdorferi Gene vlsE Involves Cassette-Specific, Segmental Gene Conversion , 1998, Infection and Immunity.

[33]  S. Norris,et al.  Kinetics and In Vivo Induction of Genetic Variation of vlsE in Borrelia burgdorferi , 1998, Infection and Immunity.

[34]  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.

[35]  J. Radolf,et al.  A new animal model for studying Lyme disease spirochetes in a mammalian host-adapted state. , 1998, The Journal of clinical investigation.

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

[37]  J. Bono,et al.  The Borrelia burgdorferi circular plasmid cp26: conservation of plasmid structure and targeted inactivation of the ospC gene , 1997, Molecular microbiology.

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

[39]  R. C. Johnson,et al.  Correlation of plasmids with infectivity of Borrelia burgdorferi sensu stricto type strain B31 , 1996, Infection and immunity.

[40]  Ying Zhang,et al.  Mutations in pncA, a gene encoding pyrazinamidase/nicotinamidase, cause resistance to the antituberculous drug pyrazinamide in tubercle bacillus , 1996, Nature Medicine.

[41]  R. Frothingham,et al.  Identification, cloning, and expression of the Escherichia coli pyrazinamidase and nicotinamidase gene, pncA , 1996, Antimicrobial agents and chemotherapy.

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

[43]  S. Norris,et al.  High- and low-infectivity phenotypes of clonal populations of in vitro-cultured Borrelia burgdorferi , 1995, Infection and immunity.

[44]  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.

[45]  P. Rosa,et al.  Plasmid location of Borrelia purine biosynthesis gene homologs , 1994, Journal of bacteriology.

[46]  Jean Côté,et al.  Lyme Disease , 1991, International journal of dermatology.

[47]  J. Roth,et al.  Activity of the nicotinamide mononucleotide transport system is regulated in Salmonella typhimurium , 1991, Journal of bacteriology.

[48]  T. Schwan,et al.  Analysis of supercoiled circular plasmids in infectious and non-infectious Borrelia burgdorferi. , 1990, Microbial pathogenesis.

[49]  Samuel L. Groseclose,et al.  Summary of Notifiable Diseases, United States. , 1997 .

[50]  R. Miller,et al.  One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

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

[52]  R. C. Johnson,et al.  Infection of Syrian hamsters with Lyme disease spirochetes , 1984, Journal of clinical microbiology.

[53]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[54]  J. Roth,et al.  6-Aminonicotinamide-resistant mutants of Salmonella typhimurium , 1983, Journal of bacteriology.

[55]  L. Reed,et al.  A SIMPLE METHOD OF ESTIMATING FIFTY PER CENT ENDPOINTS , 1938 .

[56]  L. J. Reed A simple method of estimating fifty per cent end-points. , 1938 .