Peptidoglycan Endopeptidase PBP7 Facilitates the Recruitment of FtsN to the Divisome and Promotes Peptidoglycan Synthesis in Escherichia coli.
Escherichia coli
PBP7
divisome
penicillin‐binding proteins
peptidoglycan hydrolases
peptidoglycan synthesis
Journal
Molecular microbiology
ISSN: 1365-2958
Titre abrégé: Mol Microbiol
Pays: England
ID NLM: 8712028
Informations de publication
Date de publication:
30 Sep 2024
30 Sep 2024
Historique:
revised:
07
09
2024
received:
18
03
2024
accepted:
10
09
2024
medline:
30
9
2024
pubmed:
30
9
2024
entrez:
30
9
2024
Statut:
aheadofprint
Résumé
Escherichia coli has many periplasmic hydrolases to degrade and modify peptidoglycan (PG). However, the redundancy of eight PG endopeptidases makes it challenging to define specific roles to individual enzymes. Therefore, the cellular role of PBP7 (encoded by pbpG) is not clearly defined. In this work, we show that PBP7 localizes in the lateral cell envelope and at midcell. The C-terminal α-helix of PBP7 is crucial for midcell localization but not for its activity, which is dispensable for this localization. Additionally, midcell localization of PBP7 relies on the assembly of FtsZ up to FtsN in the divisome, and on the activity of PBP3. PBP7 was found to affect the assembly timing of FtsZ and FtsN in the divisome. The absence of PBP7 slows down the assembly of FtsN at midcell. The ΔpbpG mutant exhibited a weaker incorporation of the fluorescent D-amino acid HADA, reporting on transpeptidase activity, compared to wild-type cells. This could indicate reduced PG synthesis at the septum of the ΔpbpG strain, explaining the slower accumulation of FtsN and suggesting that endopeptidase-mediated PG cleavage may be a rate-limiting step for septal PG synthesis.
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : China Scholarship Council fellowship
ID : 201804910650
Organisme : UKRI Future Leaders Fellowship
ID : MR/V027204/1
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/W005557/1
Pays : United Kingdom
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/W013630/1
Pays : United Kingdom
Organisme : Springboard award
ID : SBF005/1112
Informations de copyright
© 2024 The Author(s). Molecular Microbiology published by John Wiley & Sons Ltd.
Références
Aarsman, M. E. G., A. Piette, C. Fraipont, T. M. F. Vinkenvleugel, M. Nguyen‐Distèche, and T. den Blaauwen. 2005. “Maturation of the Escherichia coli Divisome Occurs in Two Steps.” Molecular Microbiology 55: 1631–1645.
Alcorlo, M., D. A. Dik, S. De Benedetti, et al. 2019. “Structural Basis of Denuded Glycan Recognition by SPOR Domains in Bacterial Cell Division.” Nature Communications 10: 5567.
Arends, S. J., K. Williams, R. J. Scott, S. Rolong, D. L. Popham, and D. S. Weiss. 2010. “Discovery and Characterization of Three New Escherichia coli Septal Ring Proteins That Contain a SPOR Domain: DamX, DedD, and RlpA.” Journal of Bacteriology 192: 242–255.
Bertsche, U., T. Kast, B. Wolf, et al. 2006. “Interaction Between Two Murein (Peptidoglycan) Synthases, PBP3 and PBP1B, in Escherichia coli.” Molecular Microbiology 61: 675–690.
Bi, E. F., and J. Lutkenhaus. 1991. “FtsZ Ring Structure Associated With Division in Escherichia coli.” Nature 354: 161–164.
Buddelmeijer, N., M. Aarsman, and T. den Blaauwen. 2013. “Immunolabeling of Proteins in Situ in Escherichia coli K12 Strains.” Bio‐Protocol 3: e852.
Burman, L. G., and J. T. Park. 1984. “Molecular Model for Elongation of the Murein Sacculus of Escherichia coli.” Proceedings of the National Academy of Sciences of the United States of America 81: 1844–1848.
Chen, J. C., and J. Beckwith. 2001. “FtsQ, FtsL and FtsI Require FtsK, but Not FtsN, for Co‐Localization With FtsZ During Escherichia coli Cell Division.” Molecular Microbiology 42: 395–413.
Cho, H., C. N. Wivagg, M. Kapoor, et al. 2016. “Bacterial Cell Wall Biogenesis Is Mediated by SEDS and PBP Polymerase Families Functioning Semi‐Autonomously.” Nature Microbiology 1: 16172.
Chodisetti, P. K., and M. Reddy. 2019. “Peptidoglycan Hydrolase of an Unusual Cross‐Link Cleavage Specificity Contributes to Bacterial Cell Wall Synthesis.” Proceedings of the National Academy of Sciences of the United States of America 116: 7825–7830.
Datsenko, K. A., and B. L. Wanner. 2000. “One‐Step Inactivation of Chromosomal Genes in Escherichia coli K‐12 Using PCR Products.” Proceedings of the National Academy of Sciences of the United States of America 97: 6640–6645.
Davies, T. A., M. G. Page, W. Shang, T. Andrew, M. Kania, and K. Bush. 2007. “Binding of Ceftobiprole and Comparators to the Penicillin‐Binding Proteins of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus pneumoniae.” Antimicrobial Agents and Chemotherapy 51: 2621–2624.
den Blaauwen, T., M. A. de Pedro, M. Nguyen‐Disteche, and J. A. Ayala. 2008. “Morphogenesis of Rod‐Shaped Sacculi.” FEMS Microbiology Reviews 32: 321–344.
den Blaauwen, T., L. W. Hamoen, and P. A. Levin. 2017. “The Divisome at 25: The Road Ahead.” Current Opinion in Microbiology 36: 85–94.
Du, S., W. Henke, S. Pichoff, and J. Lutkenhaus. 2019. “How FtsEX Localizes to the Z Ring and Interacts With FtsA to Regulate Cell Division.” Molecular Microbiology 112: 881–895.
Ducret, A., E. M. Quardokus, and Y. V. Brun. 2016. “MicrobeJ, a Tool for High Throughput Bacterial Cell Detection and Quantitative Analysis.” Nature Microbiology 1: 16077.
Egan, A. J., and W. Vollmer. 2013. “The Physiology of Bacterial Cell Division.” Annals of the New York Academy of Sciences 1277: 8–28.
Gerding, M. A., B. Liu, F. O. Bendezu, C. A. Hale, T. G. Bernhardt, and P. A. de Boer. 2009. “Self‐Enhanced Accumulation of FtsN at Division Sites and Roles for Other Proteins With a SPOR Domain (DamX, DedD, and RlpA) in Escherichia coli Cell Constriction.” Journal of Bacteriology 191: 7383–7401.
Gibson, D. G., L. Young, R. Y. Chuang, J. C. Venter, C. A. Hutchison 3rd, and H. O. Smith. 2009. “Enzymatic Assembly of DNA Molecules up to Several Hundred Kilobases.” Nature Methods 6: 343–345.
Glauner, B., J. V. Höltje, and U. Schwarz. 1988. “The Composition of the Murein of Escherichia coli.” Journal of Biological Chemistry 263: 10088–10095.
Goehring, N. W., M. D. Gonzalez, and J. Beckwith. 2006. “Premature Targeting of Cell Division Proteins to Midcell Reveals Hierarchies of Protein Interactions Involved in Divisome Assembly.” Molecular Microbiology 61: 33–45.
Goodell, E. W. 1985. “Recycling of Murein by Escherichia coli.” Journal of Bacteriology 163: 305–310.
Goodell, E. W., and U. Schwarz. 1985. “Release of Cell Wall Peptides Into Culture Medium by Exponentially Growing Escherichia coli.” Journal of Bacteriology 162: 391–397.
Gray, A. N., A. J. F. Egan, I. L. Van't Veer, et al. 2015. “Coordination of Peptidoglycan Synthesis and Outer Membrane Constriction During Escherichia coli Cell Division.” eLife 4: e07118.
Harz, H., K. Burgdorf, and J. V. Höltje. 1990. “Isolation and Separation of the Glycan Strands From Murein of Escherichia coli by Reversed‐Phase High‐Performance Liquid Chromatography.” Analytical Biochemistry 190: 120–128.
Heidrich, C., M. F. Templin, A. Ursinus, et al. 2001. “Involvement of N‐Acetylmuramyl‐l‐Alanine Amidases in Cell Separation and Antibiotic‐Induced Autolysis of Escherichia coli.” Molecular Microbiology 41: 167–178.
Heidrich, C., A. Ursinus, J. Berger, H. Schwarz, and J. V. Holtje. 2002. “Effects of Multiple Deletions of Murein Hydrolases on Viability, Septum Cleavage, and Sensitivity to Large Toxic Molecules in Escherichia coli.” Journal of Bacteriology 184: 6093–6099.
Höltje, J. V. 1998. “Growth of the Stress‐Bearing and Shape‐Maintaining Murein Sacculus of Escherichia coli.” Microbiology and Molecular Biology Reviews 62: 181–203.
Jeon, W. J., and H. Cho. 2022. “A Cell Wall Hydrolase MepH Is Negatively Regulated by Proteolysis Involving Prc and NlpI in Escherichia coli.” Frontiers in Microbiology 13: 878049.
Jumper, J., R. Evans, A. Pritzel, et al. 2021. “Highly Accurate Protein Structure Prediction With AlphaFold.” Nature 596: 583–589.
Kang, K. N., and J. M. Boll. 2022. “PBP1A Directly Interacts With the Divisome Complex to Promote Septal Peptidoglycan Synthesis in Acinetobacter baumannii.” Journal of Bacteriology 204: e0023922.
Koppelman, C. M., M. E. Aarsman, J. Postmus, et al. 2004. “R174 of Escherichia coli FtsZ Is Involved in Membrane Interaction and Protofilament Bundling, and Is Essential for Cell Division.” Molecular Microbiology 51: 645–657.
Korat, B., H. Mottl, and W. Keck. 1991. “Penicillin‐Binding Protein 4 of Escherichia coli: Molecular Cloning of the dacB Gene, Controlled Overexpression, and Alterations in Murein Composition.” Molecular Microbiology 1991, no. 5: 675–684.
Leclercq, S., A. Derouaux, S. Olatunji, et al. 2017. “Interplay Between Penicillin‐Binding Proteins and SEDS Proteins Promotes Bacterial Cell Wall Synthesis.” Scientific Reports 7: 43306.
Liu, X., J. Biboy, E. Consoli, W. Vollmer, and T. den Blaauwen. 2020. “MreC and MreD Balance the Interaction Between the Elongasome Proteins PBP2 and RodA.” PLoS Genetics 16: e1009276.
Lyu, Z., A. Yahashiri, X. Yang, et al. 2022. “FtsN Maintains Active Septal Cell Wall Synthesis by Forming a Processive Complex With the Septum‐Specific Peptidoglycan Synthases in E. coli.” Nature Communications 13: 5751.
Macheboeuf, P., C. Contreras‐Martel, V. Job, O. Dideberg, and A. Dessen. 2006. “Penicillin Binding Proteins: Key Players in Bacterial Cell Cycle and Drug Resistance Processes.” FEMS Microbiology Reviews 30: 673–691.
Magnet, S., L. Dubost, A. Marie, M. Arthur, and L. Gutmann. 2008. “Identification of Thel,d‐Transpeptidases for Peptidoglycan Cross‐Linking in Escherichia coli.” Journal of Bacteriology 190: 4782–4785.
Mainardi, J. L., R. Villet, T. D. Bugg, C. Mayer, and M. Arthur. 2008. “Evolution of Peptidoglycan Biosynthesis Under the Selective Pressure of Antibiotics in Gram‐Positive Bacteria.” FEMS Microbiology Reviews 32: 386–408.
Mamou, G., F. Corona, R. Cohen‐Khait, et al. 2022. “Peptidoglycan Maturation Controls Outer Membrane Protein Assembly.” Nature 606: 953–959.
Marrec‐Fairley, M., A. Piette, X. Gallet, et al. 2000. “Differential Functionalities of Amphiphilic Peptide Segments of the Cell‐Septation Penicillin‐Binding Protein 3 of Escherichia coli.” Molecular Microbiology 37: 1019–1031.
Matsuhashi, M., S. Tamaki, S. J. Curtis, and J. L. Strominger. 1979. “Mutational Evidence for Identity of Penicillin‐Binding Protein 5 in Escherichia coli With the Major D‐Alanine Carboxypeptidase IA Activity.” Journal of Bacteriology 137: 644–647.
Mertens, L. M. Y., and T. den Blaauwen. 2022. “Optimising Expression of the Large Dynamic Range FRET Pair mNeonGreen and Superfolder mTurquoise2(Ox) for Use in the Escherichia coli Cytoplasm.” Scientific Reports 12: 17977.
Mueller, E. A., C. S. Westfall, and P. A. Levin. 2020. “pH‐Dependent Activation of Cytokinesis Modulates Escherichia coli Cell Size.” PLoS Genetics 16: e1008685.
Müller, P., C. Ewers, U. Bertsche, et al. 2007. “The Essential Cell Division Protein FtsN Interacts With the Murein (Peptidoglycan) Synthase PBP1B in Escherichia coli*.” Journal of Biological Chemistry 282: 36394–36402.
Navarro, P. P., A. Vettiger, V. Y. Ananda, et al. 2022. “Cell Wall Synthesis and Remodelling Dynamics Determine Division Site Architecture and Cell Shape in Escherichia coli.” Nature Microbiology 7: 1621–1634.
Nicola, G., S. Peddi, M. Stefanova, R. A. Nicholas, W. G. Gutheil, and C. Davies. 2005. “Crystal Structure of Escherichia coli Penicillin‐Binding Protein 5 Bound to a Tripeptide Boronic Acid Inhibitor: A Role for Ser‐110 in Deacylation.” Biochemistry 44: 8207–8217.
Park, J. T., and T. Uehara. 2008. “How Bacteria Consume Their Own Exoskeletons (Turnover and Recycling of Cell Wall Peptidoglycan).” Microbiology and Molecular Biology Reviews 72: 211–227.
Park, S. H., Y. J. Kim, H. B. Lee, Y. J. Seok, and C. R. Lee. 2020. “Genetic Evidence for Distinct Functions of Peptidoglycan Endopeptidases in Escherichia coli.” Frontiers in Microbiology 11: 565767.
Pazos, M., K. Peters, M. Casanova, et al. 2018. “Z‐Ring Membrane Anchors Associate With Cell Wall Synthases to Initiate Bacterial Cell Division.” Nature Communications 9: 5090.
Peters, N. T., T. Dinh, and T. G. Bernhardt. 2011. “A Fail‐Safe Mechanism in the Septal Ring Assembly Pathway Generated by the Sequential Recruitment of Cell Separation Amidases and Their Activators.” Journal of Bacteriology 93: 4973–4983.
Peters, N. T., C. Morlot, D. C. Yang, T. Uehara, T. Vernet, and T. G. Bernhardt. 2013. “Structure‐Function Analysis of the LytM Domain of EnvC, an Activator of Cell Wall Remodelling at the Escherichia coli Division Site.” Molecular Microbiology 89: 690–701.
Pisabarro, A. G., M. A. de Pedro, and D. Vázquez. 1985. “Structural Modifications in the Peptidoglycan of Escherichia coli Associated With Changes in the State of Growth of the Culture.” Journal of Bacteriology 161: 238–242.
Pogliano, J., K. Pogliano, D. S. Weiss, R. Losick, and J. Beckwith. 1997. “Inactivation of FtsI Inhibits Constriction of the FtsZ Cytokinetic Ring and Delays the Assembly of FtsZ Rings at Potential Division Sites.” Proceedings of the National Academy of Sciences of the United States of America 94: 559–564.
Priyadarshini, R., M. A. de Pedro, and K. D. Young. 2007. “Role of Peptidoglycan Amidases in the Development and Morphology of the Division Septum in Escherichia coli.” Journal of Bacteriology 189: 5334–5347.
Priyadarshini, R., D. L. Popham, and K. D. Young. 2006. “Daughter Cell Separation by Penicillin‐Binding Proteins and Peptidoglycan Amidases in Escherichia coli.” Journal of Bacteriology 188: 5345–5355.
Rohs, P. D. A., J. Buss, S. I. Sim, et al. 2018. “A Central Role for PBP2 in the Activation of Peptidoglycan Polymerization by the Bacterial Cell Elongation Machinery.” PLoS Genetics 14: e1007726.
Romeis, T., and J. V. Höltje. 1994. “Specific Interaction of Penicillin‐Binding Proteins 3 and 7/8 With Soluble Lytic Transglycosylase in Escherichia coli.” Journal of Biological Chemistry 269: 21603–21607.
Sauvage, E., F. Kerff, M. Terrak, J. A. Ayala, and P. Charlier. 2008. “The Penicillin‐Binding Proteins: Structure and Role in Peptidoglycan Biosynthesis.” FEMS Microbiology Reviews 32: 234–258.
Singh, S. K., L. SaiSree, R. N. Amrutha, and M. Reddy. 2012. “Three Redundant Murein Endopeptidases Catalyse an Essential Cleavage Step in Peptidoglycan Synthesis of Escherichia coli K12.” Molecular Microbiology 86: 1036–1051.
Sjodt, M., K. Brock, G. Dobihal, et al. 2018. “Structure of the Peptidoglycan Polymerase RodA Resolved by Evolutionary Coupling Analysis.” Nature 556: 118–121.
Sykes, R. B., D. P. Bonner, K. Bush, and N. H. Georgopapadakou. 1982. “Azthreonam (SQ 26,776), a Synthetic Monobactam Specifically Active Against Aerobic Gram‐Negative Bacteria.” Antimicrobial Agents and Chemotherapy 21: 85–92.
Taguchi, A., M. A. Welsh, L. S. Marmont, et al. 2019. “FtsW Is a Peptidoglycan Polymerase That Is Functional Only in Complex With Its Cognate Penicillin‐Binding Protein.” Nature Microbiology 4: 587–594.
Thomason, L. C., N. Costantino, and D. L. Court. 2007. “E. coli Genome Manipulation by P1 Transduction.” Current Protocols in Molecular Biology 79: 1.17.1–1.17.8.
Ting, S. Y., D. E. Bosch, S. M. Mangiameli, et al. 2018. “Bifunctional Immunity Proteins Protect Bacteria Against FtsZ‐Targeting ADP‐Ribosylating Toxins.” Cell 175: 1380–1392.e14.
Tsang, M. J., A. A. Yakhnina, and T. G. Bernhardt. 2017. “NlpD Links Cell Wall Remodeling and Outer Membrane Invagination During Cytokinesis in Escherichia coli.” PLoS Genetics 13: e1006888.
Ursinus, A., F. van den Ent, S. Brechtel, et al. 2004. “Murein (Peptidoglycan) Binding Property of the Essential Cell Division Protein FtsN From Escherichia coli.” Journal of Bacteriology 186: 6728–6737.
Van der Ploeg, R., S. T. Goudelis, and T. Den Blaauwen. 2015. “Validation of FRET Assay for the Screening of Growth Inhibitors of Escherichia coli Reveals Elongasome Assembly Dynamics.” International Journal of Molecular Sciences 16: 17637–17654.
van Heijenoort, J. 2011. “Peptidoglycan Hydrolases of Escherichia coli.” Microbiology and Molecular Biology Reviews 75: 636–663.
Verheul, J., A. Lodge, H. C. L. Yau, et al. 2022. “Early Midcell Localization of Escherichia coli PBP4 Supports the Function of Peptidoglycan Amidases.” PLoS Genetics 18: e1010222.
Vischer, N. O., J. Verheul, M. Postma, et al. 2015. “Cell Age Dependent Concentration of Escherichia coli Divisome Proteins Analyzed With ImageJ and ObjectJ.” Frontiers in Microbiology 6: 586.
Voedts, H., D. Dorchêne, A. Lodge, W. Vollmer, M. Arthur, and J.‐E. Hugonnet. 2021. “Role of Endopeptidases in Peptidoglycan Synthesis Mediated by Alternative Cross‐Linking Enzymes in Escherichia coli.” EMBO Journal 40: e108126.
Vollmer, W., D. Blanot, and M. A. de Pedro. 2008. “Peptidoglycan Structure and Architecture.” FEMS Microbiology Reviews 32: 149–167.
Vollmer, W., B. Joris, P. Charlier, and S. Foster. 2008. “Bacterial Peptidoglycan (Murein) Hydrolases.” FEMS Microbiology Reviews 32: 259–286.
Wachi, M., M. Doi, Y. Okada, and M. Matsuhashi. 1989. “New mre Genes mreC and mreD, Responsible for Formation of the Rod Shape of Escherichia coli Cells.” Journal of Bacteriology 171: 6511–6516.
Yakhnina, A. A., and T. G. Bernhardt. 2020. “The Tol‐Pal System Is Required for Peptidoglycan‐Cleaving Enzymes to Complete Bacterial Cell Division.” Proceedings of the National Academy of Sciences of the United States of America 117: 6777–6783.
Zhang, W., Q. Shi, S. O. Meroueh, S. B. Vakulenko, and S. Mobashery. 2007. “Catalytic Mechanism of Penicillin‐Binding Protein 5 of Escherichia coli.” Biochemistry 46: 10113–10121.
Zhao, G., T. I. Meier, S. D. Kahl, K. R. Gee, and L. C. Blaszczak. 1999. “BOCILLIN FL, a Sensitive and Commercially Available Reagent for Detection of Penicillin‐Binding Proteins.” Antimicrobial Agents and Chemotherapy 43: 1124–1128.