Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

ABSTRACT ftsZ is essential for cell division in many microorganisms. In Escherichia coli and Bacillus subtilis , FtsZ plays a role in ring formation at the leading edge of the cell division septum. An ftsZ homologue is present in the Borrelia burgdorferi genome ( ftsZ Bbu ). Its gene product (FtsZ Bbu ) is strongly homologous to other bacterial FtsZ proteins, but its function has not been established. Because loss-of-function mutants of ftsZ Bbu might be lethal, the tetR/tetO system was adapted for regulated control of this gene in B. burgdorferi . Sixty-two nucleotides of an ftsZ Bbu antisense DNA sequence under the control of a tetracycline-responsive modified hybrid borrelial promoter were cloned into pKFSS1. This construct was electroporated into a B. burgdorferi host strain carrying a chromosomally located tetR under the control of the B. burgdorferi flaB promoter. After induction by anhydrotetracycline, expression of antisense ftsZ RNA resulted in generation of filamentous B. burgdorferi that were unable to divide and grew more slowly than uninduced cells. To determine whether FtsZ Bbu could interfere with the function of E. coli FtsZ, ftsZ Bbu was amplified from chromosomal DNA and placed under the control of the tetracycline-regulated hybrid promoter. After introduction of the construct into E. coli and induction with anhydrotetracycline, overexpression of ftsZ Bbu generated a filamentous phenotype. This suggested interference of ftsZ Bbu with E. coli FtsZ function and confirmed the role of ftsZ Bbu in cell division. This is the first report of the generation of a B. burgdorferi conditional lethal mutant equivalent by tetracycline-controlled expression of antisense RNA.

Bibliography

Dubytska, L., Godfrey, H. P., & Cabello, F. C. (2006). Borrelia burgdorferi ftsZ Plays a Role in Cell Division. Journal of Bacteriology, 188(5), 1969–1978.

Authors 3
  1. Lydia Dubytska (first)
  2. Henry P. Godfrey (additional)
  3. Felipe C. Cabello (additional)
References 59 Referenced 9
  1. Akins, D. R., S. F. Porcella, T. G. Popova, D. Shevchenko, S. I. Baker, M. Li, M. V. Norgard, and J. D. Radolf. 1995. Evidence for in vivo but not in vitro expression of a Borrelia burgdorferi outer surface protein F (OspF) homologue. Mol. Microbiol.18:507-520. (10.1111/j.1365-2958.1995.mmi_18030507.x) / Mol. Microbiol. (1995)
  2. 10.1046/j.1365-2958.2003.03537.x
  3. Baron, U., and H. Bujard. 2000. Tet repressor-based system for regulated gene expression in eukaryotic cells: principles and advances. Methods Enzymol.327:401-421. (10.1016/S0076-6879(00)27292-3) / Methods Enzymol. (2000)
  4. Beall, B., and J. Lutkenhaus. 1991. FtsZ in Bacillus subtilis is required for vegetative septation and for asymmetric septation during sporulation. Genes Dev.5:447-455. (10.1101/gad.5.3.447) / Genes Dev. (1991)
  5. Berens, C., and W. Hillen. 2004. Gene regulation by tetracyclines. Genet. Eng. N. Y.26:255-277. / Genet. Eng. N. Y. (2004)
  6. Bi, E. F., and J. Lutkenhaus. 1991. FtsZ ring structure associated with division in Escherichia coli. Nature354:161-164. (10.1038/354161a0) / Nature (1991)
  7. 10.1128/IAI.72.8.4864-4867.2004
  8. Bramhill, D. 1997. Bacterial cell division. Annu. Rev. Cell Dev. Biol.13:395-424. (10.1146/annurev.cellbio.13.1.395) / Annu. Rev. Cell Dev. Biol. (1997)
  9. 10.1128/IAI.73.8.4972-4981.2005
  10. Burgdorfer, W., S. F. Hayes, and J. L. Benach. 1988. Development of Borrelia burgdorferi in ixodid tick vectors. Ann. N. Y. Acad. Sci.539:172-179. (10.1111/j.1749-6632.1988.tb31851.x) / Ann. N. Y. Acad. Sci. (1988)
  11. Cabello, F. C., M. L. Sartakova, and E. Y. Dobrikova. 2001. Genetic manipulation of spirochetes: light at the end of the tunnel. Trends Microbiol.9:245-248. (10.1016/S0966-842X(01)02046-7) / Trends Microbiol. (2001)
  12. 10.1128/jb.173.11.3500-3506.1991
  13. 10.1128/jb.176.1.130-136.1994
  14. Dewar, S. J., and R. Dorazi. 2000. Control of division gene expression in Escherichia coli. FEMS Microbiol. Lett.187:1-7. (10.1111/j.1574-6968.2000.tb09127.x) / FEMS Microbiol. Lett. (2000)
  15. Ehrt, S., X. V. Guo, C. M. Hickey, M. Ryou, M. Monteleone, L. W. Riley, and D. Schnappinger. 2005. Controlling gene expression in mycobacteria with anhydrotetracycline and Tet repressor. Nucleic Acids Res.33:e21. (10.1093/nar/gni013) / Nucleic Acids Res. (2005)
  16. Erickson, H. P. 1995. FtsZ, a prokaryotic homolog of tubulin? Cell80:367-370. (10.1016/0092-8674(95)90486-7) / Cell (1995)
  17. 10.1128/JB.185.22.6723-6727.2003
  18. 10.1038/37551
  19. Gossen, M., and H. Bujard. 1992. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc. Natl. Acad. Sci. USA89:5547-5551. (10.1073/pnas.89.12.5547) / Proc. Natl. Acad. Sci. USA (1992)
  20. Gossen, M., and H. Bujard. 1993. Anhydrotetracycline, a novel effector for tetracycline controlled gene expression systems in eukaryotic cells. Nucleic Acids Res.21:4411-4412. (10.1093/nar/21.18.4411) / Nucleic Acids Res. (1993)
  21. Gossen, M., and H. Bujard. 2002. Studying gene function in eukaryotes by conditional gene inactivation. Annu. Rev. Genet.36:153-173. (10.1146/annurev.genet.36.041002.120114) / Annu. Rev. Genet. (2002)
  22. Grantcharova, N., W. Ubhayasekera, S. L. Mowbray, J. R. McCormick, and K. Flardh. 2003. A missense mutation in ftsZ differentially affects vegetative and developmentally controlled cell division in Streptomyces coelicolor A3(2). Mol. Microbiol.47:645-656. (10.1046/j.1365-2958.2003.03334.x) / Mol. Microbiol. (2003)
  23. 10.1128/JB.186.9.2673-2681.2004
  24. Horiuchi, T., H. Maki, and M. Sekiguchi. 1981. Conditional lethality of Escherichia coli strains carrying dnaE and dnaQ mutations. Mol. Gen. Genet.181:24-28. (10.1007/BF00339000) / Mol. Gen. Genet. (1981)
  25. 10.1128/JB.183.3.1078-1084.2001
  26. 10.1128/JB.181.21.6585-6590.1999
  27. Ji, Y., G. Woodnutt, M. Rosenberg, and M. K. Burnham. 2002. Identification of essential genes in Staphylococcus aureus using inducible antisense RNA. Methods Enzymol.358:123-128. (10.1016/S0076-6879(02)58084-8) / Methods Enzymol. (2002)
  28. 10.1128/AEM.71.2.728-733.2005
  29. Knee, R., and P. R. Murphy. 1997. Regulation of gene expression by natural antisense RNA transcripts. Neurochem. Int.31:379-392. (10.1016/S0197-0186(96)00108-8) / Neurochem. Int. (1997)
  30. Kukekova, A. V., A. Y. Malinin, J. A. Ayala, and S. N. Borchsenius. 1999. Characterization of Acholeplasma laidlawii ftsZ gene and its gene product. Biochem. Biophys. Res. Commun.262:44-49. (10.1006/bbrc.1999.1135) / Biochem. Biophys. Res. Commun. (1999)
  31. Lee, L. K., and C. M. Roth. 2003. Antisense technology in molecular and cellular bioengineering. Curr. Opin. Biotechnol.14:505-511. (10.1016/S0958-1669(03)00115-0) / Curr. Opin. Biotechnol. (2003)
  32. Lutz, R., and H. Bujard. 1997. Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucleic Acids Res.25:1203-1210. (10.1093/nar/25.6.1203) / Nucleic Acids Res. (1997)
  33. 10.1128/JB.181.24.7531-7544.1999
  34. 10.1128/jb.173.18.5822-5830.1991
  35. Mirochnitchenko, O., and M. Inouye. 2000. Antisense RNA and DNA in Escherichia coli. Methods Enzymol.313:467-485. (10.1016/S0076-6879(00)13030-7) / Methods Enzymol. (2000)
  36. Momynaliev, K. T., O. V. Smirnova, V. N. Lazyrev, T. A. Akopian, V. V. Chelysheva, J. A. Ayala, A. N. Simankova, S. N. Borchsenius, and V. M. Govorun. 2002. Characterization of the Mycoplasma hominis ftsZ gene and its sequence variability in mycoplasma clinical isolates. Biochem. Biophys. Res. Commun.293:155-162. (10.1016/S0006-291X(02)00184-5) / Biochem. Biophys. Res. Commun. (2002)
  37. Mukherjee, A., K. Dai, and J. Lutkenhaus. 1993. Escherichia coli cell division protein FtsZ is a guanine nucleotide binding protein. Proc. Natl. Acad. Sci. USA90:1053-1057. (10.1073/pnas.90.3.1053) / Proc. Natl. Acad. Sci. USA (1993)
  38. 10.1128/JB.180.19.5020-5029.1998
  39. Nogales, E., K. H. Downing, L. A. Amos, and J. Lowe. 1998. Tubulin and FtsZ form a distinct family of GTPases. Nat. Struct. Biol.5:451-458. (10.1038/nsb0698-451) / Nat. Struct. Biol. (1998)
  40. 10.1128/JB.181.4.1348-1351.1999
  41. Old, I. G., J. MacDougall, G. Saint, I., and B. E. Davidson. 1992. Mapping of genes on the linear chromosome of the bacterium Borrelia burgdorferi: possible locations for its origin of replication. FEMS Microbiol. Lett.78:245-250. / FEMS Microbiol. Lett. (1992)
  42. 10.1128/JB.186.24.8472-8477.2004
  43. Pfaffl, M. W. 2001. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res.29:e45. (10.1093/nar/29.9.e45) / Nucleic Acids Res. (2001)
  44. 10.1128/IAI.70.4.2029-2038.2002
  45. 10.1038/nrmicro1086
  46. Sambrook J. and D. W. Russell. 2001. Molecular cloning: a laboratory manual. 2nd ed. Cold Spring Harbor Laboratory Press Cold Spring Harbor N.Y.
  47. Schwedock, J., J. R. McCormick, E. R. Angert, J. R. Nodwell, and R. Losick. 1997. Assembly of the cell division protein FtsZ into ladder-like structures in the aerial hyphae of Streptomyces coelicolor. Mol. Microbiol.25:847-858. (10.1111/j.1365-2958.1997.mmi507.x) / Mol. Microbiol. (1997)
  48. Shevchuk, N. A., A. V. Bryksin, Y. A. Nusinovich, F. C. Cabello, M. Sutherland, and S. Ladisch. 2004. Construction of long DNA molecules using long PCR-based fusion of several fragments simultaneously. Nucleic Acids Res.32:e19. (10.1093/nar/gnh014) / Nucleic Acids Res. (2004)
  49. 10.1128/IAI.72.4.2280-2287.2004
  50. Stieger, M., B. Wohlgensinger, M. Kamber, R. Lutz, and W. Keck. 1999. Integrational plasmids for the tetracycline-regulated expression of genes in Streptococcus pneumoniae. Gene226:243-251. (10.1016/S0378-1119(98)00561-7) / Gene (1999)
  51. 10.1128/JB.186.12.3951-3959.2004
  52. 10.1128/JB.184.2.452-458.2002
  53. Trieu-Cuot, P., C. Carlier, C. Poyart-Salmeron, and P. Courvalin. 1991. An integrative vector exploiting the transposition properties of Tn1545 for insertional mutagenesis and cloning of genes from gram-positive bacteria. Gene106:21-27. (10.1016/0378-1119(91)90561-O) / Gene (1991)
  54. Urlinger, S., U. Baron, M. Thellmann, M. T. Hasan, H. Bujard, and W. Hillen. 2000. Exploring the sequence space for tetracycline-dependent transcriptional activators: novel mutations yield expanded range and sensitivity. Proc. Natl. Acad. Sci. USA97:7963-7968. (10.1073/pnas.130192197) / Proc. Natl. Acad. Sci. USA (2000)
  55. Wang, Y., B. D. Jones, and Y. V. Brun. 2001. A set of ftsZ mutants blocked at different stages of cell division in Caulobacter.Mol. Microbiol.40:347-360. (10.1046/j.1365-2958.2001.02395.x) / Mol. Microbiol. (2001)
  56. Ward, J. E., Jr., and J. Lutkenhaus. 1985. Overproduction of FtsZ induces minicell formation in Escherichia coli. Cell42:941-949. (10.1016/0092-8674(85)90290-9) / Cell (1985)
  57. Weiss, D. S. 2004. Bacterial cell division and the septal ring. Mol. Microbiol.54:588-597. (10.1111/j.1365-2958.2004.04283.x) / Mol. Microbiol. (2004)
  58. Zhang, L., F. Fan, L. M. Palmer, M. A. Lonetto, C. Petit, L. L. Voelker, A. St. John, B. Bankosky, M. Rosenberg, and D. McDevitt. 2000. Regulated gene expression in Staphylococcus aureus for identifying conditional lethal phenotypes and antibiotic mode of action. Gene255:297-305. (10.1016/S0378-1119(00)00325-5) / Gene (2000)
  59. Zhao, Y., R. W. Hammond, I. M. Lee, B. A. Roe, S. Lin, and R. E. Davis. 2004. Cell division gene cluster in Spiroplasma kunkelii: functional characterization of ftsZ and the first report of ftsA in mollicutes. DNA Cell Biol.23:127-134. (10.1089/104454904322759948) / DNA Cell Biol. (2004)
Dates
Type When
Created 19 years, 6 months ago (Feb. 15, 2006, 9:38 p.m.)
Deposited 4 years ago (July 29, 2021, 1:32 p.m.)
Indexed 1 year, 10 months ago (Sept. 28, 2023, 3:41 p.m.)
Issued 19 years, 5 months ago (March 1, 2006)
Published 19 years, 5 months ago (March 1, 2006)
Published Print 19 years, 5 months ago (March 1, 2006)
Funders 0

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@article{Dubytska_2006, title={Borrelia burgdorferi ftsZ Plays a Role in Cell Division}, volume={188}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.188.5.1969-1978.2006}, DOI={10.1128/jb.188.5.1969-1978.2006}, number={5}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Dubytska, Lydia and Godfrey, Henry P. and Cabello, Felipe C.}, year={2006}, month=mar, pages={1969–1978} }