[en] The murein (peptidoglycan) sacculus is an essential polymer embedded in the bacterial envelope. The Escherichia coli class B penicillin-binding protein (PBP) 3 is a murein transpeptidase and essential for cell division. In an affinity chromatography experiment, the bifunctional transglycosylase-transpeptidase murein synthase PBP1B was retained by PBP3-sepharose when a membrane fraction of E. coli was applied. The direct protein-protein interaction between purified PBP3 and PBP1B was characterized in vitro by surface plasmon resonance. The interaction was confirmed in vivo employing two different methods: by a bacterial two-hybrid system, and by cross-linking/co-immunoprecipitation. In the bacterial two-hybrid system, a truncated PBP3 comprising the N-terminal 56 amino acids interacted with PBP1B. Both synthases could be cross-linked in vivo in wild-type cells and in cells lacking FtsW or FtsN. PBP1B localized diffusely and in foci at the septation site and also at the side wall. Statistical analysis of the immunofluorescence signals revealed that the localization of PBP1B at the septation site depended on the physical presence of PBP3, but not on the activity of PBP3. These studies have demonstrated, for the first time, a direct interaction between a class B PBP (PBP3) and a class A PBP (PBP1B) in vitro and in vivo, indicating that different murein synthases might act in concert to enlarge the murein sacculus during cell division.
Aarsman, M.E., Piette, A., Fraipont, C., Vinkenvleugel, T.M., Nguyen-Disteche, M., and den Blaauwen, T. (2005) Maturation of the Escherichia coli divisome occurs in two steps. Mol Microbiol 55: 1631-1645.
Alaedini, A., and Day, R.A. (1999) Identification of two penicillin-binding multienzyme complexes in Haemophilus influenzae. Biochem Biophys Res Commun 264: 191-195.
Arbeloa, A., Segal, H., Hugonnet, J.E., Josseaume, N., Dubost, L., Brouard, J.P., et al. (2004) Role of class A penicillin-binding proteins in PBP5-mediated beta-lactam resistance in Enterococcus faecalis. J Bacteriol 186: 1221-1228.
Bertsche, U., Breukink, E., Kast, T., and Vollmer, W. (2005) In vitro murein (peptidoglycan) synthesis by dimers of the bifunctional transglycosylase-transpeptidase PBP1B from Escherichia coli. J Biol Chem 280: 38096-38101.
den Blaauwen, T., Wientjes, F.B., Kolk, A.H., Spratt, B.G., and Nanninga, N. (1989) Preparation and characterization of monoclonal antibodies against native membrane-bound penicillin-binding protein 1B of Escherichia coli. J Bacteriol 171: 1394-1401.
den Blaauwen, T., Pas, E., Edelman, A., Spratt, B.G., and Nanninga, N. (1990a) Mapping of conformational epitopes of monoclonal antibodies against Escherichia coli penicillin-binding protein 1B (PBP 1B) by means of hybrid protein analysis: implications for the tertiary structure of PBP 1B. J Bacteriol 172: 7284-7288.
den Blaauwen, T., Aarsman, M., and Nanninga, N. (1990b) Interaction of monoclonal antibodies with the enzymatic domains of penicillin-binding protein 1b of Escherichia coli. J Bacteriol 172: 63-70.
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., Vischer, N.O., 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.
Broome-Smith, J.K., Hedge, P.J., and Spratt, B.G. (1985) Production of thiol-penicillin-binding protein 3 of Escherichia coli using a two primer method of site-directed mutagenesis. EMBO J 4: 231-235.
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.
Casabadan, M.J., and Cohen, S.N. (1980) Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol 138: 179-207.
Charpentier, X., Chalut, C., Remy, M.H., and Masson, J.M. (2002) Penicillin-binding proteins 1a and 1b form independent dimers in Escherichia coli. J Bacteriol 184: 3749-3752.
Chen, J.C., and Beckwith, J. (2001) FtsQ, FtsL and FtsI require FtsK, but not FtsN, for co-localization with FtsZ during Escherichia coli cell division. Mol Microbiol 42: 395-413.
Corbin, B.D., Geissler, B., Sadasivam, M., and Margolin, W. (2004) Z-ring-independent interaction between a subdomain of FtsA and late septation proteins as revealed by a polar recruitment assay. J Bacteriol 186: 7736-7744.
Dai, K., and Lutkenhaus, J. (1992) The proper ratio of FtsZ to FtsA is required for cell division to occur in Escherichia coli. J Bacteriol 174: 6145-6151.
Dargis, M., and Malouin, F. (1994) Use of biotinylated beta-lactams and chemiluminescence for study and purification of penicillin-binding proteins in bacteria. Antimicrob Agents Chemother 38: 973-980.
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.
Divakaruni, A.V., Ogorzalek Loo, R.R., Xie, Y., Loo, J.A., and Gober, J.W. (2005) The cell-shape protein MreC interacts with Extracytoplasmic proteins including cell wall assembly complexes in Caulobacter crescentus. Proc Natl Acad Sci USA 102: 18602-18607.
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.
Eberhardt, C., Kuerschner, L., and Weiss, D.S. (2003) Probing the catalytic activity of a cell division-specific transpeptidase in vivo with beta-lactams. J Bacteriol 185: 3726-3734.
Edelman, A., Bowler, L., Broome-Smith, J.K., and Spratt, B.G. (1987) Use of β-lactamase fusion vector to investigate the organization of the penicillin-binding protein 1B in the cytoplasmic membrane of Escherichia coli. Mol Microbiol 1: 101-106.
Figge, R.M., Divakaruni, A.V., and Gober, J.W. (2004) MreB, the cell shape-determining bacterial actin homologue, co-ordinates cell wall morphogenesis in Caulobacter crescentus. Mol Microbiol 51: 1321-1332.
Fishov, I., Zaritsky, A., and Grover, N.B. (1995) On microbial states of growth. Mol Microbiol 15: 789-794.
Fuda, C., Hesek, D., Lee, M., Morio, K., Nowak, T., and Mobashery, S. (2005) Activation for catalysis of penicillin-binding protein 2a from methicillin-resistant Staphylococcus aureus by bacterial cell wall. J Am Chem Soc 127: 2056-2057.
Garcia del Portillo, F., and de Pedro, M.A. (1990) Differential effect of mutational impairment of penicillin-binding proteins 1A and 1B on Escherichia coli strains harboring thermosensitive mutations in the cell division genes ftsA, ftsQ, ftsZ, and pbpB. J Bacteriol 172: 5863-5870.
Garcia del Portillo, F., de Pedro, M.A., and Ayala, J.A. (1991) Identification of a new mutation in Escherichia coli that suppresses a pbpB (Ts) phenotype in the presence of penicillin-binding protein 1B. FEMS Microbiol Lett 68: 7-13.
Goehring, N.W., and Beckwith, J. (2005) Diverse paths to midcell: assembly of the bacterial cell division machinery. Curr Biol 15: R514-R526.
Goffin, C., and Ghuysen, J.M. (1998) Multimodular penicillin-binding proteins: an enigmatic family of orthologs and paralogs. Microbiol Mol Biol Rev 62: 1079-1093.
Goffin, C., Fraipont, C., Ayala, J., Terrak, M., Nguyen-Disteche, 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 confers cell septation activity on the folded structure. J Bacteriol 178: 5402-5409.
Höltje, J.V. (1993) 'Three for one' - A simple growth mechanism that guarantees a precise copy of the thin, rod-shaped murein sacculus of Escherichia coli. In Bacterial Growth and Lysis-Metabolism and Structure of the Bacterial Sacculus. de Pedro, M.A., Höltje, J.V., and Löffelhardt, W. (eds). London: Plenum Press, pp. 419-426.
Höltje, J.V. (1998) Growth of the stress-bearing and shape-maintaining murein sacculus of Escherichia coli. Microbiol Mol Biol Rev 62: 181-203.
Huls, P.G., Vischer, N.O., and Woldringh, C.L. (1999) Delayed nucleoid segregation in Escherichia coli. Mol Microbiol 33: 959-970.
Jobling, M.G., and Holmes, R.K. (2000) Identification of motifs in cholera toxin A1 polypeptide that are required for its interaction with human ADP-ribosylation factor 6 in a bacterial two-hybrid system. Proc Natl Acad Sci USA 97: 14662-14667.
Karimova, G., Pidoux, J., Ullmann, A., and Ladant, D. (1998) A bacterial two-hybrid system based on a reconstituted signal transduction pathway. Proc Natl Acad Sci USA 95: 5752-5756.
Karimova, G., Dautin, N., and Ladant, D. (2005) Interaction network among Escherichia coli membrane proteins involved in cell division as revealed by bacterial two-hybrid analysis. J Bacteriol 187: 2233-2243.
Kato, J., Suzuki, H., and Hirota, Y. (1984) Overlapping of the coding regions for alpha and gamma components of penicillin-binding protein 1b in Escherichia coli. Mol Gen Genet 196: 449-457.
Kato, J., Suzuki, H., and Hirota, Y. (1985) Dispensability of either penicillin-binding protein-1a or -1b involved in the essential process for cell elongation in Escherichia coli. Mol Gen Genet 200: 272-277.
Koppelman, C.M., Aarsman, M.E., 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.
Kraus, W., and Höltje, J.-V. (1987) Two distinct transpeptidation reactions during murein synthesis in Escherichia coli. J Bacteriol 169: 3099-3103.
Lutgenberg, B.J., Meijers, J., Peters, R., van der Hoek, P., and van Alphen, L. (1975) Electrophoretic resolution of the 'major outer membrane protein' of Escherichia coli K12 into four bands. FEBS Lett 58: 254-258.
Macheboeuf, P., Di Guilmi, A.M., Job, V., Vernet, T., Dideberg, O., and Dessen, A. (2005) Active site restructuring regulates ligand recognition in class A penicillin-binding proteins. Proc Natl Acad Sci USA 102: 577-582.
Marrec-Fairley, M., Piette, A., Gallet, X., Brasseur, R., Hara, H., Fraipont, C., et al. (2000) Differential functionalities of amphiphilic peptide segments of the cell-septation penicillin-binding protein 3 of Escherichia coli. Mol Microbiol 37: 1019-1031.
Matsuhashi, M., Wachi, M., and Ishino, F. (1990) Machinery for cell growth and division: penicillin-binding proteins and other proteins. Res Microbiol 141: 89-103.
Meisel, U., Höltje, J.V., and Vollmer, W. (2003) Overproduction of inactive variants of the murein synthase PBP1B causes lysis in Escherichia coli. J Bacteriol 185: 5342-5348.
Mercer, K.L., and Weiss, D.S. (2002) The Escherichia coli cell division protein FtsW is required to recruit its cognate transpeptidase, FtsI (PBP3), to the division site. J Bacteriol 184: 904-912.
Nguyen-Disteche, 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., Piette, A., et al. (2004) Functional analysis of the cell division protein FtsW of Escherichia coli. J Bacteriol 186: 8370-8379.
de Pedro, M.A., Quintela, J.C., Höltje, J.V., and Schwarz, H. (1997) Murein segregation in Escherichia coli. J Bacteriol 179: 2823-2834.
Piette, A., Fraipont, C., den Blaauwen, T., Aarsman, M.E., Pastoret, S., and Nguyen-Disteche, M. (2004) Structural determinants required to target penicillin-binding protein 3 to the septum of Escherichia coli. J Bacteriol 186: 6110-6117.
Pinho, M.G., and Errington, J. (2005) Recruitment of penicillin-binding protein PBP2 to the division site of Staphylococcus aureus is dependent on its transpeptidation substrates. Mol Microbiol 55: 799-807.
Pinho, M.G., de Lencastre, H., and Tomasz, A. (2001) An acquired and a native penicillin-binding protein cooperate in building the cell wall of drug-resistant staphylococci. Proc Natl Acad Sci USA 98: 10886-10891.
Pisabarro, A.G., Prats, R., Vaquez, D., and Rodriguez-Tebar, A. (1986) Activity of penicillin-binding protein 3 from Escherichia coli. J Bacteriol 168: 199-206.
Romeis, T., and Höltje, J.V. (1994) Specific interaction of penicillin-binding proteins 3 and 7/8 with soluble lytic transglycosylase in Escherichia coli. J Biol Chem 269: 21603-21607.
Scheffers, D.J., and Errington, J. (2004) PBP1 is a component of the Bacillus subtilis cell division machinery. J Bacteriol 186: 5153-5156.
Scheffers, D.J., Jones, L.J., and Errington, J. (2004) Several distinct localization patterns for penicillin-binding proteins in Bacillus subtilis. Mol Microbiol 51: 749-764.
Spratt, B.G. (1975) Distinct penicillin binding proteins involved in the division, elongation, and shape of Escherichia coli K12. Proc Natl Acad Sci USA 72: 2999-3003.
Sykes, R.B., and Bonner, D.P. (1985) Discovery and development of the monobactams. Rev Infect Dis 7 (Suppl. 4): S579-S593.
Tamaki, S., Nakajima, S., and Matsuhashi, M. (1977) Thermosensitive mutation in Escherichia coli simultaneously causing defects in penicillin-binding protein-1Bs and in enzyme activity for peptidoglycan synthesis in vitro. Proc Natl Acad Sci USA 74: 5472-5476.
Taschner, P.E., Verest, J.G., and Woldringh, C.L. (1987) Genetic and morphological characterization of ftsB and nrdB mutants of Escherichia coli. J Bacteriol 169: 19-25.
Taschner, P.E.M., Ypenburg, N., Spratt, B.G., and Woldringh, C.L. (1988) An amino acid substitution in penicillin-binding protein 3 creates pointed polar caps in Escherichia coli. J Bacteriol 170: 4828-4837.
Terrak, M., Ghosh, T.K., van Heijenoort, J., Van Beeumen, J., Lampilas, M., Aszodi, J., et al. (1999) The catalytic, glycosyl transferase and acyl transferase modules of the cell wall peptidoglycan-polymerizing penicillin-binding protein 1b of Escherichia coli. Mol Microbiol 34: 350-364.
Vollmer, W., von Rechenberg, M., and Höltje, J.V. (1999) Demonstration of molecular interactions between the murein polymerase PBP1B, the lytic transglycosylase MltA, and the scaffolding protein MipA of Escherichia coli. J Biol Chem 274: 6726-6734.
Wang, X., Possoz, C., and Sherratt, D.J. (2005) Dancing around the divisome: asymmetric chromosome segregation in Escherichia coli. Genes Dev 19: 2367-2377.
Weidel, W., and Pelzer, H. (1964) Bagshaped macromolecules - a new outlook on bacterial cell walls. Adv Enzymol 26: 193-232.
Weiss, D.S., Pogliano, K., Carson, M., Guzman, L.M., Fraipont, C., Nguyen-Disteche, M., et al. (1997) Localization of the Escherichia coli cell division protein FtsI (PBP3) to the division site and cell pole. Mol Microbiol 25: 671-681.
Weiss, D.S., Chen, J.C., Ghigo, J.M., Boyd, D., and Beckwith, J. (1999) Localization of FtsI (PBP3) to the septal ring requires its membrane anchor, the Z ring, FtsA, FtsQ, and FtsL. J Bacteriol 181: 508-520.
Wissel, M.C., and Weiss, D.S. (2004) Genetic analysis of the cell division protein FtsI (PBP3): amino acid substitutions that impair septal localization of FtsI and recruitment of FtsN. J Bacteriol 186: 490-502.
Zijderveld, C.A., Aarsman, M.E., den Blaauwen, T., and Nanninga, N. (1991) Penicillin-binding protein 1B of Escherichia coli exists in dimeric forms. J Bacteriol 173: 5740-5746.