The genes involved in division site selection in were identified using raw data from the genome project and are part of a cluster of 27 genes. When gonococcal genes were heterologously expressed as a cluster in , minicells and filaments were produced, indicating that gonococcal genes disrupted cell division in other genera. The insertional inactivation of the gene of CH811 resulted in a strain (CSRC1) with decreased viability and grossly abnormal cell division as observed by phase-contrast and electron microscopy analysis. Western blot analysis of CSRC1 confirmed that MinC was not produced. Complementation of CSRC1 by integrating a –6×His tag fusion at the locus by homologous recombination restored viability and 19 times wild-type levels of MinC expression. This slight increase of expression caused a small percentage of the complemented cells to divide aberrantly. This suggested that the 6×His tag has partially affected the stability of MinC, or that the chromosomal position of is critical to its regulation. Comparison of MinC proteins from different bacteria showed a homologous region corresponding to residues 135–230 with five conserved amino acids. Overexpression of MinC in wild-type cells induced filamentation and an mutant was successfully complemented with . Therefore, the evidence indicates that MinC from acts as a cell-division inhibitor and that its role is essential in maintaining proper division in cocci.


Article metrics loading...

Loading full text...

Full text loading...



  1. Adler, H. I., Fisher, W. D., Cohen, A. & Hardigree, A. A.(1967). Miniature Escherichia coli cells deficient in DNA. Proc Natl Acad Sci U S A 57, 321-326.[CrossRef] [Google Scholar]
  2. Ayala, J. A., Garrido, T., de Pedro, M. A. & Vicente, M.(1994). Molecular biology of bacterial septation. In Bacterial Cell Wall , pp. 73-102. Edited by J. M. Ghuysen & R. Hakenbeck. Amsterdam:Elsevier.
  3. Beveridge, T. J., Popkin, T. J. & Cole, R. M.(1994). Electron microscopy. In Methods for General and Molecular Microbiology , pp. 44-70. Edited by P. Gerhardt, R. G. E. Murray, W. A. Woods & N. R. Krieg. Washington, DC:American Society for Microbiology.
  4. Bi, E. & Lutkenhaus, J.(1993). Cell division inhibitors SulA and MinCD prevent formation of the FtsZ ring. J Bacteriol 175, 1118-1125. [Google Scholar]
  5. de Boer, P. & Crossley, R.(1989). A division inhibitor and a topological specificity factor coded for by the minicell locus determine proper placement of the division septum in Escherichia coli. Cell 56, 641-649.[CrossRef] [Google Scholar]
  6. de Boer, P., Crossley, R. & Rothfield, L.(1988). Isolation and properties of minB, a complex genetic locus involved in correct placement of the division site in Escherichia coli. J Bacteriol 170, 2106-2112. [Google Scholar]
  7. de Boer, P., Cook, W. & Rothfield, L.(1990). Bacterial cell division. Annu Rev Genet 24, 249-274.[CrossRef] [Google Scholar]
  8. de Boer, P., Crossley, R. E., Hand, A. R. & Rothfield, L.(1991). The MinD protein is a membrane ATPase required for the correct placement of the Escherichia coli division site. EMBO J 10, 4371-4380. [Google Scholar]
  9. de Boer, P., Crossley, R. & Rothfield, L.(1992). Roles of MinC and MinD in the site-specific septation block mediated by the MinCDE system of Escherichia coli. J Bacteriol 174, 63-70. [Google Scholar]
  10. Cha, J. & Stewart, G.(1997). The divIVA minicell locus of B. subtilis. J Bacteriol 179, 1671-1683. [Google Scholar]
  11. Edwards, D. & Errington, J.(1997). The Bacillus subtilis DivIVA protein targets to the division septum and controls the site specificity of cell division. Mol Microbiol 25, 905-915. [Google Scholar]
  12. Fitz-James, P.(1964). Thin sections of dividing Neisseria gonorrhoeae. J Bacteriol 87, 1477-1482. [Google Scholar]
  13. Francis, F., Ramirez-Arcos, S., Salimnia, H., Victor, C. & Dillon, J. R.(2000). Organization and transcription of the division cell wall (dcw) cluster in Neisseria gonorrhoeae. Gene 251, 141-151.[CrossRef] [Google Scholar]
  14. Gaikwad, A., Babbarwal, V., Pant, V. & Mukherjee, S. K.(2000). Pea chloroplast FtsZ can form multimers and correct the thermosensitive defect of an Escherichia coli ftsZ mutant. Mol Gen Genet 263, 213-221.[CrossRef] [Google Scholar]
  15. Goodman, S. D. & Scocca, J. J.(1988). Identification and arrangement of the DNA sequences recognized in specific transformation of Neisseria gonorrhoeae. Proc Natl Acad Sci U S A 85, 6982-6986.[CrossRef] [Google Scholar]
  16. Hebert, G. A., Pelham, P. L. & Pittman, B.(1973). Determination of the optimal ammonium sulfate concentration for the fractionation of rabbit, sheep, horse and goat antisera. Appl Microbiol 25, 26-36. [Google Scholar]
  17. Hu, Z. & Lutkenhaus, J.(1999). Topological regulation of cell division in Escherichia coli involved rapid pole to pole oscillation of the cell division inhibitor MinC under the control of MinD and MinE. Mol Microbiol 34, 82-90.[CrossRef] [Google Scholar]
  18. Hu, Z. & Lutkenhaus, J.(2000). Analysis of MinC reveals two independent domains involved in interaction with MinD and FtsZ. J Bacteriol 182, 3965-3971.[CrossRef] [Google Scholar]
  19. Hu, Z., Mukherjee, A., Pichoff, S. & Lutkenhaus, J.(1999). The MinC component of the division site selection system in Escherichia coli interacts with FtsZ to prevent polymerization. Proc Natl Acad Sci U S A 96, 14819-14824.[CrossRef] [Google Scholar]
  20. Huang, J., Cao, C. & Lutkenhaus, J.(1996). Interaction between FtsZ and inhibitors of cell division. J Bacteriol 178, 5080-5085. [Google Scholar]
  21. Janik, A., Juni, E. & Heym, G. A.(1976). Genetic transformation as a tool for detection of Neisseria gonorrhoeae. J Clin Microbiol 4, 71-81. [Google Scholar]
  22. Justice, S. S., Garcia-Lara, J. & Rothfield, L. I.(2000). Cell division inhibitors SulA and MinC/MinD block septum formation at different steps in the assembly of the Escherichia coli division machinery. Mol Microbiol 37, 410-423.[CrossRef] [Google Scholar]
  23. King, G. F., Rowland, S. L., Pan, B., Mackay, J. P., Mullen, G. P. & Rothfield, L. I.(1999). The dimerization and topological specificity functions of MinE reside in a structurally autonomous C-terminal domain. Mol Microbiol 31, 1161-1169.[CrossRef] [Google Scholar]
  24. Koch, A.(1985). How bacteria grow and divide in spite of internal hydrostatic pressure. Can J Microbiol 31, 1071-1084.[CrossRef] [Google Scholar]
  25. Koomey, M.(1998). Competence for natural transformation in Neisseria gonorrhoeae: a model system for studies of horizontal gene transfer. APMIS Suppl 84, 56-61. [Google Scholar]
  26. Lee, S. J., Gray, M. C., Guo, L., Sebo, P. & Hewlett, E. L.(1999). Epitope mapping of monoclonal antibodies against Bordetella pertussis adenylate cyclase toxin. Infect Immun 67, 2090-2095. [Google Scholar]
  27. Li, J. & Rosen, B. P.(2000). The linker peptide of the ArsA ATPase. Mol Microbiol 35, 361-367.[CrossRef] [Google Scholar]
  28. Lutkenhaus, J. & Addinall, S.(1997). Bacterial cell division and the Z ring. Annu Rev Biochem 66, 93-116.[CrossRef] [Google Scholar]
  29. Margolin, W.(2000). Self-assembling GTPases caught in the middle. Curr Biol 10, R328-R330.[CrossRef] [Google Scholar]
  30. Marston, A. L. & Errington, J.(1999). Selection of the midcell division site in Bacillus subtilis through MinD-dependent polar localization and activation of MinC. Mol Microbiol 33, 84-96.[CrossRef] [Google Scholar]
  31. Marston, A. L., Thomaides, H. B., Edwards, D. H., Sharpe, M. E. & Errington, J.(1998). Polar localization of the MinD protein of Bacillus subtilis and its role in selection of the mid-cell division site. Genes Dev 12, 3419-3430.[CrossRef] [Google Scholar]
  32. Miloso, M., Limauro, D., Alifano, P., Rivellini, F., Lavitola, A., Gulleta, E. & Bruni, C. B.(1999). Characterization of the rho genes of Neisseria gonorrhoeae and Salmonella typhimurium. J Bacteriol 175, 8030-8037. [Google Scholar]
  33. Miroux, B. & Walker, J. E.(1996). Overproduction of proteins in Escherichia coli: mutant hosts that allow synthesis of some membrane proteins and globular proteins at high levels. J Mol Biol 260, 289-298.[CrossRef] [Google Scholar]
  34. Nanninga, N.(1998). Morphogenesis of Escherichia coli. Microbiol Mol Biol Rev 62, 110-129. [Google Scholar]
  35. Pagotto, F., Aman, A. T., Ng, L.-K., Yeung, K.-H., Brett, M. & Dillon, J. R.(2000). Sequence analysis of the family of penicillinase-producing plasmids of Neisseria gonorrhoeae. Plasmid 43, 24-34.[CrossRef] [Google Scholar]
  36. Pichoff, S., Vollrath, B., Touriol, C. & Bouché, J.(1995). Deletion analysis of gene minE which encodes the topological specificity factor of cell division in Escherichia coli. Mol Microbiol 18, 321-329.[CrossRef] [Google Scholar]
  37. Platt, T. & Richardson, J. P.(1992).Escherichia coli Rho factor: protein and enzyme of transcription termination. In Transcriptional Regulation , pp. 365-388. Edited by S. L. McKnight & K. R. Yamamoto. Cold Spring Harbor, NY:Cold Spring Harbor Laboratory.
  38. Raskin, D. M. & de Boer, P. A. J.(1997). The MinE ring: an FtsZ-independent cell structure required for selection of the correct division site in E. coli. Cell 91, 685-694.[CrossRef] [Google Scholar]
  39. Raskin, D. M. & de Boer, P. A. J.(1999a). Rapid pole-to-pole oscillation of a protein required for directing division to the middle of Escherichia coli. Proc Natl Acad Sci U S A 96, 4971-4976.[CrossRef] [Google Scholar]
  40. Raskin, D. M. & de Boer, P. A. J.(1999b). MinDE-dependent pole-to-pole oscillation of division inhibitor MinC in Escherichia coli. J Bacteriol 181, 6419-6424. [Google Scholar]
  41. Reeve, J. N., Mendelson, N. H., Coyne, S. I., Hallock, L. L. & Cole, R. M.(1973). Minicells of Bacillus subtilis. J Bacteriol 114, 860-873. [Google Scholar]
  42. Roe, B. A., Lin, S. P., Song, L., Yuan, X., Clifton, S., Ducey, T., Lewis, L. & Dyer, D. W. (2000). Gonococcal Genome Sequencing Project (Funded by USPHS/ NIH grant AI 38399). University of Oklahoma.
  43. Rothfield, L., Justice, S. & Garcı́a-Lara, J. (1999). Bacterial cell division. Annu Rev Genet 33, 423-448.[CrossRef] [Google Scholar]
  44. Rowland, S. L., Fu, X., Sayed, M. A., Zhang, Y., Cook, W. R. & Rothfield, L. I.(2000). Membrane redistribution of the Escherichia coli MinD protein induced by MinE. J Bacteriol 182, 613-619.[CrossRef] [Google Scholar]
  45. Saitoh, F., Kawamura, S., Yamasaki, N., Tanaka, I. & Kimura, M.(1999). Arginine-55 in the beta-arm is essential for the activity of DNA-binding protein HU from Bacillus stearothermophilus. Biosci Biotechnol Biochem 63, 2232-2235.[CrossRef] [Google Scholar]
  46. Salimnia, H., Radia, A., Bernatchez, S. & Dillon, J. R.(2000). Characterization of the ftsZ cell division gene of N. gonorrhoeae: expression and localization in E. coli and N. gonorrhoeae. Arch Microbiol 173, 10-20.[CrossRef] [Google Scholar]
  47. Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989).Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  48. Sen, M. & Rothfield, L. I.(1998). Stability of the Escherichia coli division inhibitor protein MinC requires determinants in the carboxy-terminal region of the protein. J Bacteriol 180, 175-177. [Google Scholar]
  49. Silver, L. E. & Clark, V. L.(1995). Construction of a translational lacZ fusion system to study gene regulation in Neisseria gonorrhoeae. Gene 166, 101-104.[CrossRef] [Google Scholar]
  50. Thoden, J. B., Kappock, T. J., Stubbe, J. & Holden, H. M.(1999). Three-dimensional structure of N5-carboxyaminoimidazole ribonucleotide synthetase: a member of the ATP grasp protein superfamily. Biochemistry 38, 15480-15492.[CrossRef] [Google Scholar]
  51. Varley, A. & Stewart, G.(1992). The divIVB region of the Bacillus subtilis chromosome encoded homologs of E. coli septum placement (MinCD) and cell shape (MreBCD) determinants. J Bacteriol 174, 6729-6742. [Google Scholar]
  52. Westling-Häggström, B., Elmros, T., Normark, S. & Winblad, B.(1977). Growth pattern and cell division in Neisseria gonorrhoeae. J Bacteriol 129, 333-342. [Google Scholar]
  53. Wu, J. & Filutowics, M.(1999). Hexahistidine (His6)-tag dependent protein dimerization: a cautionary tale. Acta Biochim Pol 46, 591-599. [Google Scholar]
  54. Yu, X. & Margolin, W.(1999). FtsZ ring clusters in min and partition mutants: role of both the Min system and the nucleoid in regulating FtsZ ring localization. Mol Microbiol 32, 315-326.[CrossRef] [Google Scholar]
  55. Zhang, Y., Rowland, S., King, G., Braswell, E. & Rothfield, L.(1998). The relationship between hetero-oligomer formation and function of the topological specificity domain of the Escherichia coli MinE protein. Mol Microbiol 30, 265-273.[CrossRef] [Google Scholar]
  56. Zhao, C., de Boer, P. & Rothfield, L.(1995). Proper placement of the Escherichia coli division site requires two functions that are associated with different domains of the MinE protein. Proc Natl Acad Sci U S A 92, 4313-4317.[CrossRef] [Google Scholar]

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error