Genetic analysis of the septal peptidoglycan synthase FtsWI complex supports a conserved activation mechanism for SEDS-bPBP complexes

Autoři: Ying Li aff001;  Han Gong aff001;  Rui Zhan aff001;  Shushan Ouyang aff001;  Kyung-Tae Park aff002;  Joe Lutkenhaus aff002;  Shishen Du aff001
Působiště autorů: Department of Microbiology, Hubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Wuhan University, Wuhan, HB, China aff001;  Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS, United States of America aff002
Vyšlo v časopise: Genetic analysis of the septal peptidoglycan synthase FtsWI complex supports a conserved activation mechanism for SEDS-bPBP complexes. PLoS Genet 17(4): e1009366. doi:10.1371/journal.pgen.1009366
Kategorie: Research Article


SEDS family peptidoglycan (PG) glycosyltransferases, RodA and FtsW, require their cognate transpeptidases PBP2 and FtsI (class B penicillin binding proteins) to synthesize PG along the cell cylinder and at the septum, respectively. The activities of these SEDS-bPBPs complexes are tightly regulated to ensure proper cell elongation and division. In Escherichia coli FtsN switches FtsA and FtsQLB to the active forms that synergize to stimulate FtsWI, but the exact mechanism is not well understood. Previously, we isolated an activation mutation in ftsW (M269I) that allows cell division with reduced FtsN function. To try to understand the basis for activation we isolated additional substitutions at this position and found that only the original substitution produced an active mutant whereas drastic changes resulted in an inactive mutant. In another approach we isolated suppressors of an inactive FtsL mutant and obtained FtsWE289G and FtsIK211I and found they bypassed FtsN. Epistatic analysis of these mutations and others confirmed that the FtsN-triggered activation signal goes from FtsQLB to FtsI to FtsW. Mapping these mutations, as well as others affecting the activity of FtsWI, on the RodA-PBP2 structure revealed they are located at the interaction interface between the extracellular loop 4 (ECL4) of FtsW and the pedestal domain of FtsI (PBP3). This supports a model in which the interaction between the ECL4 of SEDS proteins and the pedestal domain of their cognate bPBPs plays a critical role in the activation mechanism.

Klíčová slova:

Antibiotics – Arabinose – Cell cycle and cell division – Hyperexpression techniques – Library screening – Protein interactions – Signaling cascades – Substitution mutation


1. Du S, Lutkenhaus J. Assembly and activation of the Escherichia coli divisome. Mol Microbiol. 2017;105(2):177–87. doi: 10.1111/mmi.13696 28419603

2. Egan AJF, Errington J, Vollmer W. Regulation of peptidoglycan synthesis and remodelling. Nat Rev Microbiol. 2020;18(8):446–60. doi: 10.1038/s41579-020-0366-3 32424210

3. Taguchi A, Welsh MA, Marmont LS, Lee W, Sjodt M, Kruse AC, et al. FtsW is a peptidoglycan polymerase that is functional only in complex with its cognate penicillin-binding protein. Nat Microbiol. 2019;4(4):587–94. doi: 10.1038/s41564-018-0345-x 30692671

4. Yang X, McQuillen R, Lyu Z, Phillips-Mason P, De La Cruz A, McCausland JW, et al. A two-track model for the spatiotemporal coordination of bacterial septal cell wall synthesis revealed by single-molecule imaging of FtsW. Nat Microbiol. 2021. doi: 10.1038/s41564-020-00853-0 33495624

5. Bertsche U, Kast T, Wolf B, Fraipont C, Aarsman ME, Kannenberg K, et al. Interaction between two murein (peptidoglycan) synthases, PBP3 and PBP1B, in Escherichia coli. Mol Microbiol. 2006;61(3):675–90. doi: 10.1111/j.1365-2958.2006.05280.x 16803586

6. Muller P, Ewers C, Bertsche U, Anstett M, Kallis T, Breukink E, et al. The essential cell division protein FtsN interacts with the murein (peptidoglycan) synthase PBP1B in Escherichia coli. J Biol Chem. 2007;282(50):36394–402. doi: 10.1074/jbc.M706390200 17938168

7. Paradis-Bleau C, Markovski M, Uehara T, Lupoli TJ, Walker S, Kahne DE, et al. Lipoprotein cofactors located in the outer membrane activate bacterial cell wall polymerases. Cell. 2010;143(7):1110–20. doi: 10.1016/j.cell.2010.11.037 21183074

8. Typas A, Banzhaf M, van den Berg van Saparoea B, Verheul J, Biboy J, Nichols RJ, et al. Regulation of peptidoglycan synthesis by outer-membrane proteins. Cell. 2010;143(7):1097–109. doi: 10.1016/j.cell.2010.11.038 21183073

9. Boes A, Olatunji S, Breukink E, Terrak M. Regulation of the Peptidoglycan Polymerase Activity of PBP1b by Antagonist Actions of the Core Divisome Proteins FtsBLQ and FtsN. mBio. 2019;10(1). doi: 10.1128/mBio.01912-18 30622193

10. Boes A, Kerff F, Herman R, Touze T, Breukink E, Terrak M. The bacterial cell division protein fragment (E)FtsN binds to and activates the major peptidoglycan synthase PBP1b. J Biol Chem. 2020;295(52):18256–65. doi: 10.1074/jbc.RA120.015951 33109614

11. Ikeda M, Sato T, Wachi M, Jung HK, Ishino F, Kobayashi Y, et al. Structural similarity among Escherichia coli FtsW and RodA proteins and Bacillus subtilis SpoVE protein, which function in cell division, cell elongation, and spore formation, respectively. J Bacteriol. 1989;171(11):6375–8. doi: 10.1128/jb.171.11.6375-6378.1989 2509435

12. Meeske AJ, Riley EP, Robins WP, Uehara T, Mekalanos JJ, Kahne D, et al. SEDS proteins are a widespread family of bacterial cell wall polymerases. Nature. 2016;537(7622):634–8. doi: 10.1038/nature19331 27525505

13. Lara B, Ayala JA. Topological characterization of the essential Escherichia coli cell division protein FtsW. FEMS Microbiol Lett. 2002;216(1):23–32. doi: 10.1111/j.1574-6968.2002.tb11409.x 12423747

14. Cho H, Wivagg CN, Kapoor M, Barry Z, Rohs PDA, Suh H, et al. Bacterial cell wall biogenesis is mediated by SEDS and PBP polymerase families functioning semi-autonomously. Nat Microbiol. 2016;1:16172. doi: 10.1038/nmicrobiol.2016.172 27643381

15. Rohs PDA, Buss J, Sim SI, Squyres GR, Srisuknimit V, Smith M, et al. A central role for PBP2 in the activation of peptidoglycan polymerization by the bacterial cell elongation machinery. PLoS Genet. 2018;14(10):e1007726. doi: 10.1371/journal.pgen.1007726 30335755

16. Sjodt M, Rohs PDA, Gilman MSA, Erlandson SC, Zheng S, Green AG, et al. Structural coordination of polymerization and crosslinking by a SEDS-bPBP peptidoglycan synthase complex. Nat Microbiol. 2020;5(6):813–20. doi: 10.1038/s41564-020-0687-z 32152588

17. Shiomi D, Toyoda A, Aizu T, Ejima F, Fujiyama A, Shini T, et al. Mutations in cell elongation genes mreB, mrdA and mrdB suppress the shape defect of RodZ-deficient cells. Mol Microbiol. 2013;87(5):1029–44. doi: 10.1111/mmi.12148 23301723

18. Liu B, Persons L, Lee L, de Boer PA. Roles for both FtsA and the FtsBLQ subcomplex in FtsN-stimulated cell constriction in Escherichia coli. Mol Microbiol. 2015;95(6):945–70. doi: 10.1111/mmi.12906 25496160

19. Tsang MJ, Bernhardt TG. A role for the FtsQLB complex in cytokinetic ring activation revealed by an ftsL allele that accelerates division. Mol Microbiol. 2015;95(6):925–44. doi: 10.1111/mmi.12905 25496050

20. Du S, Pichoff S, Lutkenhaus J. FtsEX acts on FtsA to regulate divisome assembly and activity. Proc Natl Acad Sci U S A. 2016;113(34):E5052–61. doi: 10.1073/pnas.1606656113 27503875

21. Marmont LS, Bernhardt TG. A conserved subcomplex within the bacterial cytokinetic ring activates cell wall synthesis by the FtsW-FtsI synthase. Proc Natl Acad Sci U S A. 2020;117(38):23879–85. doi: 10.1073/pnas.2004598117 32907942

22. Park KT, Du S, Lutkenhaus J. Essential Role for FtsL in Activation of Septal Peptidoglycan Synthesis. mBio. 2020;11(6). doi: 10.1128/mBio.03012-20 33293384

23. Du S, Pichoff S, Lutkenhaus J. Roles of ATP Hydrolysis by FtsEX and Interaction with FtsA in Regulation of Septal Peptidoglycan Synthesis and Hydrolysis. mBio. 2020;11(4). doi: 10.1128/mBio.01247-20 32636250

24. Addinall SG, Cao C, Lutkenhaus J. FtsN, a late recruit to the septum in Escherichia coli. Mol Microbiol. 1997;25(2):303–9. doi: 10.1046/j.1365-2958.1997.4641833.x 9282742

25. Lutkenhaus J. FtsN—trigger for septation. J Bacteriol. 2009;191(24):7381–2. doi: 10.1128/JB.01100-09 19854895

26. Gerding MA, Liu B, Bendezu FO, Hale CA, Bernhardt TG, de Boer PA. 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. J Bacteriol. 2009;191(24):7383–401. doi: 10.1128/JB.00811-09 19684127

27. Tsang MJ, Bernhardt TG. Guiding divisome assembly and controlling its activity. Curr Opin Microbiol. 2015;24:60–5. doi: 10.1016/j.mib.2015.01.002 25636132

28. Bernard CS, Sadasivam M, Shiomi D, Margolin W. An altered FtsA can compensate for the loss of essential cell division protein FtsN in Escherichia coli. Mol Microbiol. 2007;64(5):1289–305. doi: 10.1111/j.1365-2958.2007.05738.x 17542921

29. Modell JW, Hopkins AC, Laub MT. A DNA damage checkpoint in Caulobacter crescentus inhibits cell division through a direct interaction with FtsW. Genes Dev. 2011;25(12):1328–43. doi: 10.1101/gad.2038911 21685367

30. Modell JW, Kambara TK, Perchuk BS, Laub MT. A DNA damage-induced, SOS-independent checkpoint regulates cell division in Caulobacter crescentus. PLoS Biol. 2014;12(10):e1001977. doi: 10.1371/journal.pbio.1001977 25350732

31. Mercer KL, Weiss DS. The Escherichia coli cell division protein FtsW is required to recruit its cognate transpeptidase, FtsI (PBP3), to the division site. J Bacteriol. 2002;184(4):904–12. doi: 10.1128/jb.184.4.904-912.2002 11807049

32. Wissel MC, Wendt JL, Mitchell CJ, Weiss DS. The transmembrane helix of the Escherichia coli division protein FtsI localizes to the septal ring. J Bacteriol. 2005;187(1):320–8. doi: 10.1128/JB.187.1.320-328.2005 15601716

33. Arends SJ, Kustusch RJ, Weiss DS. ATP-binding site lesions in FtsE impair cell division. J Bacteriol. 2009;191(12):3772–84. doi: 10.1128/JB.00179-09 19376877

34. Wissel MC, Weiss DS. Genetic analysis of the cell division protein FtsI (PBP3): amino acid substitutions that impair septal localization of FtsI and recruitment of FtsN. J Bacteriol. 2004;186(2):490–502. doi: 10.1128/jb.186.2.490-502.2004 14702319

35. Ovchinnikov S, Kinch L, Park H, Liao Y, Pei J, Kim DE, et al. Large-scale determination of previously unsolved protein structures using evolutionary information. Elife. 2015;4:e09248. doi: 10.7554/eLife.09248 26335199

36. Sjodt M, Brock K, Dobihal G, Rohs PDA, Green AG, Hopf TA, et al. Structure of the peptidoglycan polymerase RodA resolved by evolutionary coupling analysis. Nature. 2018;556(7699):118–21. doi: 10.1038/nature25985 29590088

37. Cong Q, Anishchenko I, Ovchinnikov S, Baker D. Protein interaction networks revealed by proteome coevolution. Science. 2019;365(6449):185–9. doi: 10.1126/science.aaw6718 31296772

38. Liu X, Biboy J, Consoli E, Vollmer W, den Blaauwen T. MreC and MreD balance the interaction between the elongasome proteins PBP2 and RodA. PLoS Genet. 2020;16(12):e1009276. doi: 10.1371/journal.pgen.1009276 33370261

39. Contreras-Martel C, Martins A, Ecobichon C, Trindade DM, Mattei PJ, Hicham S, et al. Molecular architecture of the PBP2-MreC core bacterial cell wall synthesis complex. Nat Commun. 2017;8(1):776. doi: 10.1038/s41467-017-00783-2 28974686

40. Sassine J, Xu M, Sidiq KR, Emmins R, Errington J, Daniel RA. Functional redundancy of division specific penicillin-binding proteins in Bacillus subtilis. Mol Microbiol. 2017;106(2):304–18. doi: 10.1111/mmi.13765 28792086

41. Reichmann NT, Tavares AC, Saraiva BM, Jousselin A, Reed P, Pereira AR, et al. SEDS-bPBP pairs direct lateral and septal peptidoglycan synthesis in Staphylococcus aureus. Nat Microbiol. 2019;4(8):1368–77. doi: 10.1038/s41564-019-0437-2 31086309

42. Cho H, Uehara T, Bernhardt TG. Beta-lactam antibiotics induce a lethal malfunctioning of the bacterial cell wall synthesis machinery. Cell. 2014;159(6):1300–11. doi: 10.1016/j.cell.2014.11.017 25480295

43. Du S, Henke W, Pichoff S, Lutkenhaus J. How FtsEX localizes to the Z ring and interacts with FtsA to regulate cell division. Mol Microbiol. 2019. doi: 10.1111/mmi.14324 31175681

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