[en] Cell division proteins FtsZ ( FtsA, ZipA, ZapA), FtsE/X, FtsK, FtsQ, FtsL/B, FtsW, PBP3, FtsN and AmiC localize at mid cell in Escherichia coli in an interdependent order as listed. To investigate whether this reflects a time dependent maturation of the divisome, the average cell age at which FtsZ, FtsQ, FtsW, PBP3 and FtsN arrive at their destination was determined by immuno- and GFP- fluorescence microscopy of steady state grown cells at a variety of growth rates. Consistently, a time delay of 14 - 21 min, depending on the growth rate, between Z-ring formation and the mid cell recruitment of proteins down stream of FtsK was found. We suggest a two-step model for bacterial division in which the Z-ring is involved in the switch from cylindrical to polar peptidoglycan synthesis, whereas the much later localizing cell division proteins are responsible for the modification of the envelope shape into that of two new poles.
Adam, M., Damblon, C., Jamin, M., Zorzi, W., Dusart, V., Galleni, M., et al. (1991) Acyltransferase activities of the high-molecular-mass essential penicillin-binding proteins. Biochem J 279: 601-604.
Bernhardt, T.G., and de Boer, P.A. (2003) The Escherichia coli amidase AmiC is a periplasmic septal ring component exported via the twin-arginine transport pathway. Mol Microbiol 48: 1171-1182.
Bi, E., and Lutkenhaus, J. (1990) Analysis of ftsZ mutations that confer resistance to the cell division inhibitor SulA (SfiA). J Bacteriol 172: 5602-5609.
Boyd, D., Weiss, D.S., Chen, J.C., and Beckwith, J. (2000) Towards single-copy gene expression systems making gene cloning physiologically relevant: lambda InCh, a simple Escherichia coli plasmid-chromosome shuttle system. J Bacteriol 182: 842-847.
Buddelmeijer, N., and Beckwith, J. (2004) A complex of the Escherichia coli cell division proteins FtsL, FtsB and FtsQ forms independently of its localization to the septal region. Mol Microbiol 52: 1315-1327.
Buddelmeijer, N., Aarsman, M.E.G., Kolk, A.H.J., Vicente, M., and Nanninga, N. (1998) Localization of cell division protein FtsQ by immunofluorescence microscopy in dividing and non-dividing cells of Escherichia coli. J Bacteriol 180: 6107-6116.
Buddelmeijer, N., Judson, N., Boyd, D., Mekalanos, J.J., and Beckwith, J. (2002) YgbQ, a cell division protein in Escherichia coli and Vibrio cholerae, localizes in codependent fashion with FtsL to the division site. Proc Natl Acad Sci USA 99: 6316-6321.
Carson, M.J., Barondess, J., and Beckwith, J. (1991) The FtsQ protein of Escherichia coli: membrane topology, abundance, and cell division phenotypes due to overproduction and insertion mutations. J Bacteriol 173: 2187-2195.
Chen, J.C., Weiss, D.S., Ghigo, J.-M., and Beckwith, J. (1999) Septal localization of FtsQ, an essential cell division protein in Escherichia coli. J Bacteriol 181: 521-530.
Cormack, B.P., Valdivia, R.H., and Falkow, S. (1996) FACS-optimized mutants of the green fluorescent protein (GFP). Genes Dev 173: 33-38.
Dai, K., Xu, Y., and Lutkenhaus, J. (1993) Cloning and characterization of ftsN, an essential cell division gene in Escherichia coli isolated as a multicopy suppressor of ftsA12 (ts). J Bacteriol 175: 3790-3797.
Dai, K., Xu, Y., and Lutkenhaus, J. (1996) Topological characterization of the essential Escherichia coli cell division protein FtsN. J Bacteriol 178: 1328-1334.
De Pedro, M.A., Quintela, J.C., Höltje, J.-V., and Schwarz, H. (1997) Murein segregation in Escherichia coli. J Bacteriol 179: 2823-2834.
Den Blaauwen, T., Aarsman, M.E.G., and Nanninga, N. (1990) Interaction of monoclonal antibodies with the enzymatic domains of penicillin binding protein 1b of Escherichia coli. J Bacteriol 172: 63-70.
Den Blaauwen, T., Buddelmeijer, N., Aarsman, M.E.G., Hameete, C.M., and Nanninga, N. (1999) Timing of the FtsZ assembly in Escherichia coli. J Bacteriol 181: 5167-5175.
Den Blaauwen, T., Lindqvist, A., Löwe, J., and Nanninga, N. (2001) Distribution of the Escherichia coli structural maintenance of chromosomes (SMC)-like protein MukB in the cell. Mol Microbiol 42: 1179-1188.
Den Blaauwen, T., Aarsman, M.E.G., Vischer, N.O.E., and Nanninga, N. (2003) Penicillin Binding protein PBP2 of Escherichia coli, localizes preferentially in the lateral wall and at mid cell in comparison with the old cell pole. Mol Microbiol 47: 539-547.
Di Berardino, M., Dijkstra, A., Stuber, D., Keck, W., and Gubler, M. (1996) The monofunctional glycosyltransferase of Escherichia coli is a member of a new class of peptidoglycan-synthesising enzymes. FEBS Lett 392: 184-188.
Di Lallo, G., Fagioli, M., Barionovi, D., Ghelardini, P., and Paolozzi, L. (2003) Use of a two-hybrid assay to study the assembly of a complex multicomponent protein machinery: bacterial septosome differentiation. Microbiology 149: 3353-3359.
Dougherty, T.J., Kennedy, K., Kessler, R.E., and Pucci, M.J. (1996) Direct quantitation of the number of individual penicillin-binding proteins per cell in Escherichia coli. J Bacteriol 178: 6110-6115.
Errington, J., Daniel, R.A., and Scheffers, D.J. (2003) Cytokinesis in bacteria. Microbiol Mol Biol Rev 67: 52-65.
Fishov, I., Zaritsky, A., and Graver, N.B. (1995) On microbial states of growth. Mol Microbiol 15: 789-794.
Fraipont, C., Adam, M., Nguyen-Distèche, M., Keck, W., van Beeumen, J., Ayala, J.A., et al. (1994) Engineering and overexpression of periplasmic forms of the penicillin-binding protein 3 of Escherichia coli. Biochem J 298: 189-195.
Fujiwara, T., Ritchie, K., Murakoshi, H., Jacobson, K., and Kusumi, A. (2002) Phospholipids undergo hop diffusion in compartmentalized cell membrane. J Cell Biol 157: 1071-1081.
Goffin, C., Fraipont, C., Ayala, J., Terak, M., Nguyen-Distèche, M., and Ghuysen, J.-M. (1996) The non-penicillin binding module of the tripartite penicillin-binding protein 3 of Escherichia coli is required for folding and/or stability of the penicillin-binding module and the membrane-anchoring module convers cell septation activity on the folded structure. J Bacteriol 178: 5402-5409.
Gueiros-Filho, F.J., and Losick, R. (2002) A widely conserved bacterial cell division protein that promotes assembly of the tubulin-like protein FtsZ. Genes Dev 16: 2544-2556.
Heidrich, C., Templin, M.F., Ursinus, A., Merdanovic, M., Berger, J., Schwarz, H., et al. (2001) Involvement of N-acetylmuramyl-L-alanine amidases in cell separation and antibiotic-induced autolysis of Escherichia coli. Mol Microbiol 41: 167-178.
Huls, P.G., Visscher, N.O.E., and Woldringh, C.L. (1999) Delayed nucleoid segregation in Escherichia coli. Mol Microbiol 33: 959-970.
Koppelman, C.M., Aarsman, M.E.G., Postmus, J., Pas, E., Muijsers, A.O., Scheffers, D.J., et al. (2004) R174 of Escherichia coli FtsZ is involved in membrane interaction and protofilament bundling, and is essential for cell division. Mol Microbiol 51: 645-657.
Lindahl, L. (1975) Intermediates and time kinetics of the in vivo assembly of Escherichia coli ribosomes. J Mol Biol 92: 15-37.
Maniatis, T., Fritsch, E.F., and Sambrook, J. (1989) Molecular Cloning: a Laboratory Manual, 2nd edn. New York: Cold Spring Harbor Laboratory.
Marec-Fairley, M., Piette, A., Gallet, X., Brasseur, R., Hara, H., Fraipont, C., et al. (2000) Differential functionalities of ampichilic peptide segments of the cell-septation penicillin-binding protein 3 of Escherichia coli. Mol Microbiol 37: 1019-1031.
Morlot, C., Zapun, A., Dideberg, O., and Vernet, T. (2003) Growth and division of Streptococcus pneumoniae: localization of the high molecular weight penicillin-binding proteins during the cell cycle. Mol Microbiol 50: 845-855.
Morlot, C., Noirclerc-Savoye, M., Zapun, A., Dideberg, O., and Vernet, T. (2004) The D,D-carboxypeptidase PBP3 organizes the division process of Streptococcus pneumoniae. Mol Microbiol 51: 1641-1648.
Nakano, Y., Yoshida, Y., Yamashita, Y., and Koga, T. (1995) Construction of a series of pACYC-derived plasmid vectors. Gene 162: 157-158.
Nanninga, N. (1991) Cell division and peptidoglycan assembly in Escherichia coli. Mol Microbiol 5: 791-795.
Nanninga, N. (1998) Morphogenesis of Escherichia coli. Microbiol Mol Biol Rev 62: 110-129.
Nguyen-Distèche, M., Fraipont, C., Buddelmeijer, N., and Nanninga, N. (1998) The structure and function of Escherichia coli penicillin-binding protein 3. Cell Mol Life Sci 54: 309-316.
Pastoret, S., Fraipont, C., den Blaauwen, T., Wolf, B., Aarsman, M.E.G., Piette, A., et al. (2004) Functional analysis of the cell division protein FtsW of Escherchia coli. J Bacteriol 186: 8370-8379.
Pichoff, S., and Lutkenhaus, J. (2002) Unique and overlapping roles for ZipA and FtsA in septal ring assembly in Escherichis coli. EMBO J 21: 685-693.
Powell, E.O. (1956) Growth rate and generation time of bacteria, with special reference to continous culture. J Gen Microbiol 15: 492-511.
Rueda, S., Vicente, M., and Mingorance, J. (2003) Concentration and assembly of the division ring proteins FtsZ, FtsA, and ZipA during the Escherichia coli cell cycle. J Bacteriol 185: 3344-3351.
Saffman, P.G., and Delbruck, M. (1975) Brownian motion in biological membranes. Proc Natl Acad Sci USA 72: 3111-3113.
Schmidt, K.L., Peterson, N.D., Kustusch, R.J., Wissel, M.C., Graham, B., Phillips, G.J., and Weiss, D.S. (2004) A predicted ABC transporter, FtsEX, is needed for cell division in Escherichia coli. J Bacteriol 186: 785-793.
Storts, D.R., and Markovitz, A. (1991) A novel rho promoter::Tn10 mutation suppresses an ftsQ1 (ts) missense mutation in an essential Escherichia coli cell division gene by a mechanism not involving polarity suppression. J Bacteriol 173: 655-663.
Taschner, P.E.M., Huls, P.G., Pas, E., and Woldringh, C.L. (1988) Division behaviour and shape changes in isogenic ftsZ, ftsQ, ftsA, pbpB, and ftsE cell division mutants of Escherichia coli during temperature shift experiments. J Bacteriol 170: 1533-1540.
Ursinus, A., van den Ent, F., Brechtel, S., de Pedro, M., Höltje, J.V., Löwe, J., and Vollmer, W. (2004) Murein (peptidoglycan) binding property of the essential cell division protein FtsN from Escherichia coli. J Bacteriol 186: 67288-66737.
Van Helvoort, J.M.L.M., and Woldringh, C.L. (1994) Nucleoid partitioning in Escherichia coli during steady-state growth and upon recovery from chloramphenicol treatment. Mol Microbiol 13: 577-583.
Varma, A., and Young, K.D. (2004) FtsZ collaborates with Penicillin Binding Proteins to generate bacterial cell shape in Escherichia coli. J Bacteriol 186: 6768-6774.
Voskuil, J.L.A., Westerbeek, C.A.M., Wu, C., Kolk, A.H.J., and Nanninga, N. (1994) Epitope mapping of Escherichia coli cell division protein FtsZ with monoclonal antibodies. J Bacteriol 176: 1886-1893.
Wang, X., Huang, J., Mukherjee, A., Cao, C., and Lutkenhaus, J. (1997) Analysis of the interaction of FtsZ with itself, GTP, and FtsA. J Bacteriol 179: 5551-5559.
Weiss, D.S., Chen, J.C., Ghigo, J.-M., Boyd, D., and Beckwith, J. (1999) Localization of Ftsl (PBP3) to the septal ring requires its membrane anchor, the Z ring, FtsA, FtsQ, and FtsL. J Bacteriol 181: 508-520.
Wientjes, F.B., and Nanninga, N. (1989) Rate and topography of peptidoglycan synthesis during cell division in Escherichia coli: concept of a leading edge. J Bacteriol 171: 3412-3419.
Wientjes, F.B., Olijhoek, A.J.M., Schwarz, U., and Nanninga, N. (1983) Laneling patterns of major penicillin-binding proteins of Escherichia coli during the cell division cycle. J Bacteriol 153: 1287-1293.
Woldringh, C.L., Valkenburg, J.A., Pas, E., Taschner, P.E.M., Huls, P., and Wientjes, F.B. (1985) Physiological and geometrical conditions for cell growth and division in Escherichia coli. Ann Inst Pasteur/Microbiol 136A: 131-138.
Woldringh, C.L., Huls, P., Pas, E., Brakenhoff, G.J., and Nanninga, N. (1987) Topography of peptidoglycan synthesis during elongation and polar cap formation in a cell division mutant of Escherichia coli MC4100. J Gen Microbiol 133: 575-586.
Yang, J.C., Van Den Ent, F., Neuhaus, D., Brevier, J., and Lowe, J. (2004) Solution structure and domain architecture of the divisome protein FtsN. Mol Microbiol 52: 651-660.