[en] The class B M1-V577 penicillin-binding protein (PBP) 3 of Escherichia coli consists of a M1-L39 membrane anchor (bearing a cytosolic tail) that is linked via a G40-S70 intervening peptide to an R71-I236 non-catalytic module (containing the conserved motifs 1-3) itself linked via motif 4 to a D237-V577 catalytic module (containing the conserved motifs 5-7 of the penicilloyl serine transferases superfamily). It has been proposed that during cell septation the peptidoglycan crosslinking activity of the acyl transferase module of PBP3 is regulated by the associated M1-I236 polypeptide itself in interaction with other components of the divisome. The fold adopted by the R71-V577 polypeptide of PBP3 has been modelled by reference to the corresponding R76-S634 polypeptide of the class B Streptococcus pneumoniae PBP2x. Based on these data and the results of site-directed mutagenesis of motifs 1-3 and of peptide segments of high amphiphilicity (identified from hydrophobic moment plots), the M1-I236 polypeptide of PBP3 appears to be precisely designed to work in the way proposed. The membrane anchor and the G40-S70 sequence (containing the G57-Q66 peptide segment) upstream from the non-catalytic module have the information ensuring that PBP3 undergoes proper insertion within the divisome at the cell septation site. Motif 1 and the I74-L82 overlapping peptide segment, motif 2 and the H160-G172 overlapping peptide segment, and the G188-D197 motif 3 are located at or close to the intermodule junction. They contain the information ensuring that PBP3 folds correctly and the acyl transferase catalytic centre adopts the active configuration. The E206-V217 peptide segment is exposed at the surface of the non-catalytic module. It has the information ensuring that PBP3 fulfils its cell septation activity within the fully complemented divisome.
Research Center/Unit :
CIP - Centre d'Ingénierie des Protéines - ULiège
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Marrec-Fairley, Monique; Université de Liège - ULiège > Centre d'Ingénierie des Protéines
Piette, André ; Université de Liège - ULiège > Centre d'Ingénierie des Protéines
Gallet, Xavier; Faculté Universitaire des Sciences Agronomiques de Gembloux - FUSAGx
Brasseur, Robert ; Faculté Universitaire des Sciences Agronomiques de Gembloux - FUSAGx
Hara, Hirochi; Saitama University Japan > Biochemistry and Molecular Biology
Fraipont, Claudine ; Université de Liège - ULiège > Centre d'Ingénierie des Protéines
Ghuysen, Jean-Marie ; Université de Liège - ULiège > Centre d'Ingénierie des Protéines
Adam M., Fraipont C., Rhazi N., Nguyen-Disteche M., Lakaye B., Frere J.M. (1997) The bimodular G57-V577 polypeptide chain of the class B penicillin-binding protein 3 of Escherichia coli catalyzes peptide bond formation from thiolesters and does not catalyze glycan chain polymerization from the lipid II intermediate. J Bacteriol 179:6005-6009.
Begg K.J., Takasuga A., Edwards D.H., Dewar S.J., Spratt B.G., Adachi H. (1990) The balance between different peptidoglycan precursors determines whether Escherichia coli cells will elongate or divide. J Bacteriol 172:6697-6703.
Bi E., Lutkenhaus J. (1990) FtsZ regulates frequency of cell division in Escherichia coli. J Bacteriol 172:2765-2768.
De Boer P.A.J., Crossley R.E., Rothfield L.I. (1992) The essential bacterial cell division protein FtsZ is a GTPase. Nature 359:254-256.
Broome-Smith J.K., Hedge P.J., 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.
Churchward G., Belin D., Nagamine Y. (1984) pSC101-derived plasmid which shows no sequence homology to other commonly used cloning vectors. Gene 31:165-171.
Degnen G.E., Cox E.C. (1974) A conditional mutator gene in Escherichia coli: Isolation, mapping and effector studies. J Bacteriol 117:477-487.
De Loof H., Rosseneu M., Brasseur R., Ruysschaert J.M. (1986) Use of hydrophobicity profiles to predict receptor binding domains on apolipoprotein E and the low density lipoprotein apolipoprotein B-E receptor. Proc Natl Acad Sci USA 83:2295-2299.
Eisenberg D. (1984) Three-dimensional structure of membrane and surface proteins. Annu Rev Biochem 53:595-623.
Eisenberg D., Weiss R.M., Terwilliger T.C. (1982) The helical hydrophobic moment: A measure of the amphiphilicity of a helix. Nature 299:371-374.
Fraipont C., Adam M., Nguyen-Disteche M., Keck W., Van Beeumen J., Ayala J. (1994) Engineering and overexpression of periplasmic forms of the penicillin-binding protein 3 of E. coli. Biochem J 298:189-195.
Goffin C., Ghuysen J.M. (1998) Multimodular penicillin-binding proteins: An enigmatic family of orthologs and paralogs. Microbiol Mol Biol Rev 62:1079-1093.
Goffin C., Ayala J.A., Nguyen-Disteche M., Ghuysen J.M. (1993) Site-directed mutagenesis of dicarboxylic acid residues of the penicillin-binding module of the Escherichia coli penicillin-binding prodein 3. FEMS Microbiol Lett 113:247-252.
Goffin C., Fraipont C., Ayala J., Terrak M., Nguyen-Disteche M., 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.
Guzman L.M., Belin D., Carson M.J., Beckwith J. (1995) Tight regulation, modulation, and high-level expression by vectors containing the arabinose P(BAD) promoter. J Bacteriol 177:4121-4130.
Guzman L.M., Weiss D.S., Beckwith J. (1997) Domain-swapping analysis of Ftsl, FtsL, and FtsQ, bitopic membrane proteins essential for cell division in Escherichia coli. J Bacteriol 179:5094-5103.
Hale C.A., De Boer P.A.J. (1997) Direct binding of FtsZ to ZipA, an essential component of the septal ring structure that mediates cell division in Escherichia coli. Cell 88:1-20.
Hara H., Yasuda S., Horiuchi K., Park J.T. (1997) A promoter for the first nine genes of the Escherichia coli mra cluster of cell division and cell envelope biosynthesis genes, including ftsI and ftsW. J Bacteriol 179:5802-5811.
Van Heijenoort J. (1996) Murein synthesis., Escherichia coli and Salmonella. Neidhardt, F.C. (ed.). Washington, DC: American Society for Microbiology Press; 1025-1034.
Higgins D.G., Sharp P.M. (1988) CLUSTAL: A package for performing multiple sequence alignment on a microcomputer. Gene 73:237-244.
Holm L., Sander C. (1991) Database algorithm for generating protein backbone and side chain co-ordinates from a C (α) trace. J Mol Biol 218:183-194.
Holm L., Sander C. (1992) Fast and simple Monte Carlo algorithm for side chain optimization in proteins. Proteins 14:213-223.
Houba-Herin N., Hara H., Inouye M., Hirota Y. (1985) Binding of penicillin to thiol-penicillin-binding protein 3 of Escherichia coli: Identification of its active site. Mol Gen Genet 201:499-504.
Ishidate K., Ursinus A., Holtje J.V., Rothfield L. (1998) Analysis of the length distribution of murein glycan strands in ftsZ and ftsI mutants of Escherichia coli. FEMS Microbiol Lett 168:71-75.
Labesse G., Colloc'h N., Pothier J., Mornon J.P. (1997) P-SEA: A new efficient assignment of secondary structure from Cα trace of proteins. Comput Appl Biosci 13:291-295.
Laskowski R.A., MacArthur M.W., Moss D.S., Thornton J.M. (1993) PROCHECK: A program to check the stereochemical quality of protein structures. J Appl Crystallogr 26:283-291.
Lowe J., Amos L.A. (1998) Crystal structure of the bacterial cell-division protein FtsZ. Nature 391:203-206.
Lutkenhaus J., Addinall S.G. (1997) A comprehensive review of the role of FtsZ in cell division in E. coli and other bacteria. Annu Rev Biochem 66:93-116.
Masui Y., Coleman J., Inouye M. (1983) Multipurpose expression cloning vehicles in Escherichia coli., Experimental Manipulation of Gene Expression. Inouye, M. (ed.). New York: Academic Press; 15-32.
Nagasawa H., Sakagami Y., Suzuki A., Suzuki H., Hara H., Hirota Y. (1989) Determination of the cleavage site involved in C-terminal processing of penicillin-binding protein 3 of Escherichia coli. J Bacteriol 171:5890-5893.
Nakagawa J., Tamaki S., Tomioka S., Matsuhashi M. (1984) Functional biosynthesis of cell wall peptidoglycan by polymorphic bifunctional polypeptides. J Biol Chem 259:13937-13946.
Nanninga N. (1998) Morphogenesis of Escherichia coli. Microbiol Mol Biol Rev 62:110-129.
Nguyen-Disteche M., Fraipont C., Buddelmeijer N., Nanninga N. (1998) The structure and function of Escherichia coli penicillin-binding protein 3. Cell Mol Life Sci 54:309-316.
Pares S., Mouz N., Petillot Y., Hakenbeck R., Dideberg O. (1996) X-ray structure of Streptococcus pneumoniae PBP2x, a primary penicillin target enzyme. Nature Struct Biol 3:284-289.
Pedersen L.B., Angert E.R., Setlow P. (1999) Septal localization of penicillin-binding protein 1 in Bacillus subtilis. J Bacteriol 181:3201-3211.
Del Portillo F.G., 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.
Poupon A., Mornon J.P. (1998) Populations of hydrophobic amino acids within protein globular domains: Identification of conserved 'topohydrophobic' positions. Proteins Struct Funct Genet 33:329-342.
Rahman M., Brasseur R. (1994) WinMGM: A fast CPK molecular graphics program for analyzing molecular structure. J Mol Graphics 12:212-218.
RayChaudhuri D., Park J.T. (1992) Escherichia coli cell division gene ftsZ encodes a novel GTP-binding protein. Nature 359:251-254.
Sali A., Blundell T.L. (1993) Comparative protein modelling by satisfaction spatial restraints. J Mol Biol 234:779-815.
Sambrook J., Fritsch E.F., Maniatis T., Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press; 1989.
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.
Taschner P.E.M., Ypenburg N., Spratt B.G., 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. (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.
Wang L., Lutkenhaus J. (1998) FtsK is an essential cell division protein that is localized to the septum and induced as part of the SOS response. Mol Microbiol 29:731-740.
Wang L., Khattar M.K., Donachie W.D., Lutkenhaus J. (1998) FtsI and FtsW are localized to the septum in Escherichia coli. J Bacteriol 180:2810-2816.
Weiss D.S., Chen J.C., Ghigo J.M., Boyd D., 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.
Wientjes F.B., Nanninga N. (1991) On the role of the high molecular weight penicillin-binding proteins in the cell cycle of Escherichia coli. Res Microbiol 142:333-344.