Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1997 Sep 1;16(17):5455–5463. doi: 10.1093/emboj/16.17.5455

Ca2+-mediated GTP-dependent dynamic assembly of bacterial cell division protein FtsZ into asters and polymer networks in vitro.

X C Yu 1, W Margolin 1
PMCID: PMC1170176  PMID: 9312004

Abstract

FtsZ, a tubulin-like GTPase that forms a dynamic ring marking the division plane of prokaryotic cells, is essential for cytokinesis. It is not known what triggers FtsZ ring assembly. In this work, we use a FtsZ-green fluorescent protein (Gfp) chimera to assay FtsZ assembly over time by using fluorescence microscopy. We show that FtsZ polymers can assemble dynamically in solution in a GTP-dependent manner. Initially, FtsZ nucleation centers grow into aster-like structures that dramatically resemble microtubule organizing centers. As assembly proceeds further, protofilament bundles emanating from different asters interconnect, mimicking the closure of the FtsZ ring in vivo. Surprisingly, millimolar levels of Ca2+ promote FtsZ dynamic assembly. FtsZ can undergo repeated GTP-dependent assembly and disassembly in solution by sequential addition and removal of Ca2+. In addition, GTP binding and hydrolysis by FtsZ are regulated by Ca2+ concentration. Although the concentration of Ca2+ required for FtsZ assembly in vitro is high, its clear and specific effect on FtsZ dynamics suggests the possibility that Ca2+ may have a role in regulating FtsZ ring assembly in the cell.

Full Text

The Full Text of this article is available as a PDF (314.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Addinall S. G., Bi E., Lutkenhaus J. FtsZ ring formation in fts mutants. J Bacteriol. 1996 Jul;178(13):3877–3884. doi: 10.1128/jb.178.13.3877-3884.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Addinall S. G., Lutkenhaus J. FtsA is localized to the septum in an FtsZ-dependent manner. J Bacteriol. 1996 Dec;178(24):7167–7172. doi: 10.1128/jb.178.24.7167-7172.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Addinall S. G., Lutkenhaus J. FtsZ-spirals and -arcs determine the shape of the invaginating septa in some mutants of Escherichia coli. Mol Microbiol. 1996 Oct;22(2):231–237. doi: 10.1046/j.1365-2958.1996.00100.x. [DOI] [PubMed] [Google Scholar]
  4. Archer J., Solomon F. Deconstructing the microtubule-organizing center. Cell. 1994 Feb 25;76(4):589–591. doi: 10.1016/0092-8674(94)90496-0. [DOI] [PubMed] [Google Scholar]
  5. Baumann P., Jackson S. P. An archaebacterial homologue of the essential eubacterial cell division protein FtsZ. Proc Natl Acad Sci U S A. 1996 Jun 25;93(13):6726–6730. doi: 10.1073/pnas.93.13.6726. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bendjennat M., Blanchard A., Loutfi M., Montagnier L., Bahraoui E. Purification and characterization of Mycoplasma penetrans Ca2+/Mg2+-dependent endonuclease. J Bacteriol. 1997 Apr;179(7):2210–2220. doi: 10.1128/jb.179.7.2210-2220.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Berkowitz S. A., Wolff J. Intrinsic calcium sensitivity of tubulin polymerization. The contributions of temperature, tubulin concentration, and associated proteins. J Biol Chem. 1981 Nov 10;256(21):11216–11223. [PubMed] [Google Scholar]
  8. Bi E. F., Lutkenhaus J. FtsZ ring structure associated with division in Escherichia coli. Nature. 1991 Nov 14;354(6349):161–164. doi: 10.1038/354161a0. [DOI] [PubMed] [Google Scholar]
  9. Bramhill D., Thompson C. M. GTP-dependent polymerization of Escherichia coli FtsZ protein to form tubules. Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):5813–5817. doi: 10.1073/pnas.91.13.5813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chang C. F., Shuman H., Somlyo A. P. Electron probe analysis, X-ray mapping, and electron energy-loss spectroscopy of calcium, magnesium, and monovalent ions in log-phase and in dividing Escherichia coli B cells. J Bacteriol. 1986 Sep;167(3):935–939. doi: 10.1128/jb.167.3.935-939.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Erickson H. P. FtsZ, a prokaryotic homolog of tubulin? Cell. 1995 Feb 10;80(3):367–370. doi: 10.1016/0092-8674(95)90486-7. [DOI] [PubMed] [Google Scholar]
  12. Erickson H. P., Taylor D. W., Taylor K. A., Bramhill D. Bacterial cell division protein FtsZ assembles into protofilament sheets and minirings, structural homologs of tubulin polymers. Proc Natl Acad Sci U S A. 1996 Jan 9;93(1):519–523. doi: 10.1073/pnas.93.1.519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gangola P., Rosen B. P. Maintenance of intracellular calcium in Escherichia coli. J Biol Chem. 1987 Sep 15;262(26):12570–12574. [PubMed] [Google Scholar]
  14. Grant J. Incidents in the Life of a Physician. Can Med Assoc J. 1916 Apr;6(4):300–304. [PMC free article] [PubMed] [Google Scholar]
  15. Hale C. A., de Boer P. A. Direct binding of FtsZ to ZipA, an essential component of the septal ring structure that mediates cell division in E. coli. Cell. 1997 Jan 24;88(2):175–185. doi: 10.1016/s0092-8674(00)81838-3. [DOI] [PubMed] [Google Scholar]
  16. Job D., Fischer E. H., Margolis R. L. Rapid disassembly of cold-stable microtubules by calmodulin. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4679–4682. doi: 10.1073/pnas.78.8.4679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Liu B., Marc J., Joshi H. C., Palevitz B. A. A gamma-tubulin-related protein associated with the microtubule arrays of higher plants in a cell cycle-dependent manner. J Cell Sci. 1993 Apr;104(Pt 4):1217–1228. doi: 10.1242/jcs.104.4.1217. [DOI] [PubMed] [Google Scholar]
  18. Ma X., Ehrhardt D. W., Margolin W. Colocalization of cell division proteins FtsZ and FtsA to cytoskeletal structures in living Escherichia coli cells by using green fluorescent protein. Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):12998–13003. doi: 10.1073/pnas.93.23.12998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Margolin W., Wang R., Kumar M. Isolation of an ftsZ homolog from the archaebacterium Halobacterium salinarium: implications for the evolution of FtsZ and tubulin. J Bacteriol. 1996 Mar;178(5):1320–1327. doi: 10.1128/jb.178.5.1320-1327.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Moritz M., Braunfeld M. B., Sedat J. W., Alberts B., Agard D. A. Microtubule nucleation by gamma-tubulin-containing rings in the centrosome. Nature. 1995 Dec 7;378(6557):638–640. doi: 10.1038/378638a0. [DOI] [PubMed] [Google Scholar]
  21. Mukherjee A., Dai K., Lutkenhaus J. Escherichia coli cell division protein FtsZ is a guanine nucleotide binding protein. Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):1053–1057. doi: 10.1073/pnas.90.3.1053. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mukherjee A., Lutkenhaus J. Guanine nucleotide-dependent assembly of FtsZ into filaments. J Bacteriol. 1994 May;176(9):2754–2758. doi: 10.1128/jb.176.9.2754-2758.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Norris V. A calcium flux at the termination of replication triggers cell division in Escherichia coli. Hypothesis. Cell Calcium. 1989 Nov-Dec;10(8):511–517. doi: 10.1016/0143-4160(89)90012-2. [DOI] [PubMed] [Google Scholar]
  24. Norris V., Grant S., Freestone P., Canvin J., Sheikh F. N., Toth I., Trinei M., Modha K., Norman R. I. Calcium signalling in bacteria. J Bacteriol. 1996 Jul;178(13):3677–3682. doi: 10.1128/jb.178.13.3677-3682.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Norris V., Seror S. J., Casaregola S., Holland I. B. A single calcium flux triggers chromosome replication, segregation and septation in bacteria: a model. J Theor Biol. 1988 Oct 7;134(3):341–350. doi: 10.1016/s0022-5193(88)80065-1. [DOI] [PubMed] [Google Scholar]
  26. Ordal G. W. Calcium ion regulates chemotactic behaviour in bacteria. Nature. 1977 Nov 3;270(5632):66–67. doi: 10.1038/270066a0. [DOI] [PubMed] [Google Scholar]
  27. Rampersaud A., Utsumi R., Delgado J., Forst S. A., Inouye M. Ca2(+)-enhanced phosphorylation of a chimeric protein kinase involved with bacterial signal transduction. J Biol Chem. 1991 Apr 25;266(12):7633–7637. [PubMed] [Google Scholar]
  28. RayChaudhuri D., Park J. T. Escherichia coli cell-division gene ftsZ encodes a novel GTP-binding protein. Nature. 1992 Sep 17;359(6392):251–254. doi: 10.1038/359251a0. [DOI] [PubMed] [Google Scholar]
  29. Rothfield L. I., Justice S. S. Bacterial cell division: the cycle of the ring. Cell. 1997 Mar 7;88(5):581–584. doi: 10.1016/s0092-8674(00)81899-1. [DOI] [PubMed] [Google Scholar]
  30. Serrano L., Valencia A., Caballero R., Avila J. Localization of the high affinity calcium-binding site on tubulin molecule. J Biol Chem. 1986 May 25;261(15):7076–7081. [PubMed] [Google Scholar]
  31. Shu H. B., Joshi H. C. Gamma-tubulin can both nucleate microtubule assembly and self-assemble into novel tubular structures in mammalian cells. J Cell Biol. 1995 Sep;130(5):1137–1147. doi: 10.1083/jcb.130.5.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shu H. B., Li Z., Palacios M. J., Li Q., Joshi H. C. A transient association of gamma-tubulin at the midbody is required for the completion of cytokinesis during the mammalian cell division. J Cell Sci. 1995 Sep;108(Pt 9):2955–2962. doi: 10.1242/jcs.108.9.2955. [DOI] [PubMed] [Google Scholar]
  33. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
  34. Solomon F. Binding sites for calcium on tubulin. Biochemistry. 1977 Feb 8;16(3):358–363. doi: 10.1021/bi00622a003. [DOI] [PubMed] [Google Scholar]
  35. Tisa L. S., Adler J. Cytoplasmic free-Ca2+ level rises with repellents and falls with attractants in Escherichia coli chemotaxis. Proc Natl Acad Sci U S A. 1995 Nov 7;92(23):10777–10781. doi: 10.1073/pnas.92.23.10777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Vallee R. B., Bloom G. S. Isolation of sea urchin egg microtubules with taxol and identification of mitotic spindle microtubule-associated proteins with monoclonal antibodies. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6259–6263. doi: 10.1073/pnas.80.20.6259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wang X., Lutkenhaus J. FtsZ ring: the eubacterial division apparatus conserved in archaebacteria. Mol Microbiol. 1996 Jul;21(2):313–319. doi: 10.1046/j.1365-2958.1996.6421360.x. [DOI] [PubMed] [Google Scholar]
  38. Wolff J., Knipling L., Sackett D. L. Charge-shielding and the "paradoxical" stimulation of tubulin polymerization by guanidine hydrochloride. Biochemistry. 1996 May 7;35(18):5910–5920. doi: 10.1021/bi9527395. [DOI] [PubMed] [Google Scholar]
  39. Zheng Y., Wong M. L., Alberts B., Mitchison T. Nucleation of microtubule assembly by a gamma-tubulin-containing ring complex. Nature. 1995 Dec 7;378(6557):578–583. doi: 10.1038/378578a0. [DOI] [PubMed] [Google Scholar]
  40. de Boer P., Crossley R., Rothfield L. The essential bacterial cell-division protein FtsZ is a GTPase. Nature. 1992 Sep 17;359(6392):254–256. doi: 10.1038/359254a0. [DOI] [PubMed] [Google Scholar]
  41. de Pereda J. M., Leynadier D., Evangelio J. A., Chacón P., Andreu J. M. Tubulin secondary structure analysis, limited proteolysis sites, and homology to FtsZ. Biochemistry. 1996 Nov 12;35(45):14203–14215. doi: 10.1021/bi961357b. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES