1887

Access Microbiology open research platform logo

Volume 6, Issue 11

Cell division cycle fluctuation of Pal concentration in Open Access

Abstract

The Tol-Pal proteins stabilize the outer membrane during cell division in many Gram-negative bacteria, including . Pal is an outer membrane lipoprotein that can bind peptidoglycan. It accumulates at the septum during division by a mobilization-and-capture mechanism. This work further substantiates and extends knowledge of Pal’s localization in using immunolabelling; this method enables the detection of endogenous proteins. The midcell localization of Pal and TolB, as seen with fluorescent protein fusions, during cell division, was confirmed. The retention of Pal in newly formed cell poles seemed to persist longer than observed with fluorescent Pal fusions. The concentration of endogenous Pal during the cell division cycle fluctuated: it decreased initially (to half the fluorescence concentration (32.1 au µm) of the maximum (64.1 au µm) reached during the cell cycle) and then increased during the second half of the cell division cycle. We probed for possible regulators and proposed two new putative regulators of Pal. By deleting the periplasmic protease, Prc decreased the total Pal abundance (to ~65% of the fluorescence concentration in WT cells) and affected its concentration fluctuation during the cell cycle. This suggests that Prc controls a cell division stage-specific regulator of Pal. Immunolabelling also supported the prediction that the small RNA MicA suppresses Pal expression (the fluorescence concentration of Pal in cells without MicA is double that of Pal in WT cells). However, the regulation by MicA occurred in a cell cycle-independent manner. All these findings urge further research on the tight regulation of the dividing cell envelope stability.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): cell division , immunolabelling , outer membrane , periplasm , protease and Tol-Pal
Funding
This study was supported by the:
  • China Scholarship Council (Award 201804910650)
    • Principle Award Recipient: XinweiLiu
  • H2020 European Research Council (Award 721484)
    • Principle Award Recipient: LaureenMertens
© 2024 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
Loading

Article metrics loading...

/content/journal/acmi/10.1099/acmi.0.000759.v3
2024-11-13
2024-12-02
Loading full text...

Full text loading...

/deliver/fulltext/acmi/6/11/acmi000759.v3.html?itemId=/content/journal/acmi/10.1099/acmi.0.000759.v3&mimeType=html&fmt=ahah

References

  1. Mertens LMY, den Blaauwen T, Liu X, Verheul J. Cell division cycle fluctuation of Pal concentration in Escherichia coli. University of Amsterdam / Amsterdam University of Applied Sciences. Figshare 2023 [View Article]
     [Google Scholar]
  2. Mamou G, Corona F, Cohen-Khait R, Housden NG, Yeung V et al. Peptidoglycan maturation controls outer membrane protein assembly. Nature 2022; 606:953–959 [View Article] [PubMed]
     [Google Scholar]
  3. Som N, Reddy M. Cross-talk between phospholipid synthesis and peptidoglycan expansion by a cell wall hydrolase. Proc Natl Acad Sci U S A 2023; 120:e2300784120 [View Article] [PubMed]
     [Google Scholar]
  4. Hummels KR, Berry SP, Li Z, Taguchi A, Min JK et al. Coordination of bacterial cell wall and outer membrane biosynthesis. Nature 2023; 615:300–304 [View Article] [PubMed]
     [Google Scholar]
  5. Egan AJF. Bacterial outer membrane constriction: outer membrane constriction. Mol Microbiol 2018; 107:676–687 [View Article]
     [Google Scholar]
  6. Szczepaniak J, Press C, Kleanthous C. The multifarious roles of Tol-Pal in Gram-negative bacteria. FEMS Microbiol Rev 2020; 44:490–506 [View Article] [PubMed]
     [Google Scholar]
  7. Tan WB, Chng S. Genetic interaction mapping highlights key roles of the Tol-Pal complex. Mol Microbiol 2022; 117:921–936 [View Article] [PubMed]
     [Google Scholar]
  8. Shrivastava R, Jiang X, Chng SS. Outer membrane lipid homeostasis via retrograde phospholipid transport in Escherichia coli: a physiological function for the Tol-Pal complex. Mol Microbiol 2017; 106:395–408
     [Google Scholar]
  9. Yakhnina AA, Bernhardt TG. The Tol-Pal system is required for peptidoglycan-cleaving enzymes to complete bacterial cell division. Proc Natl Acad Sci U S A 2020; 117:6777–6783 [View Article] [PubMed]
     [Google Scholar]
  10. Park S, Cho H. The Tol-Pal system plays an important role in maintaining cell integrity during elongation in Escherichia coli. Front Microbiol 2022; 13:891926 [View Article] [PubMed]
     [Google Scholar]
  11. Szczepaniak J, Holmes P, Rajasekar K, Kaminska R, Samsudin F et al. The lipoprotein Pal stabilises the bacterial outer membrane during constriction by a mobilisation-and-capture mechanism. Nat Commun 2020; 11:1305 [View Article] [PubMed]
     [Google Scholar]
  12. Webby MN, Williams-Jones DP, Press C, Kleanthous C. Force-generation by the trans-envelope Tol-Pal system. Front Microbiol 2022; 13:852176 [View Article] [PubMed]
     [Google Scholar]
  13. Bonsor DA, Hecht O, Vankemmelbeke M, Sharma A, Krachler AM et al. Allosteric β-propeller signalling in TolB and its manipulation by translocating colicins. EMBO J 2009; 28:2846–2857 [View Article] [PubMed]
     [Google Scholar]
  14. Parsons LM, Lin F, Orban J. Peptidoglycan recognition by Pal, an outer membrane lipoprotein. Biochemistry 2006; 45:2122–2128 [View Article] [PubMed]
     [Google Scholar]
  15. Connolley L, Szczepaniak J, Kleanthous C, Murray SM. The quantitative basis for the redistribution of immobile bacterial lipoproteins to division septa. PLoS Comput Biol 2021; 17:e1009756 [View Article] [PubMed]
     [Google Scholar]
  16. Demchick P, Koch AL. The permeability of the wall fabric of Escherichia coli and Bacillus subtilis. J Bacteriol 1996; 178:768–773 [View Article] [PubMed]
     [Google Scholar]
  17. Bouveret E, Derouiche R, Rigal A, Lloubès R, Lazdunski C et al. Peptidoglycan-associated lipoprotein-TolB interaction. J Biol Chem 1995; 270:11071–11077 [View Article]
     [Google Scholar]
  18. Clavel T, Germon P, Vianney A, Portalier R, Lazzaroni JC. TolB protein of Escherichia coli K-12 interacts with the outer membrane peptidoglycan-associated proteins Pal, Lpp and OmpA. Mol Microbiol 1998; 29:359–367 [View Article] [PubMed]
     [Google Scholar]
  19. Park JS, Lee WC, Yeo KJ, Ryu K-S, Kumarasiri M et al. Mechanism of anchoring of OmpA protein to the cell wall peptidoglycan of the gram-negative bacterial outer membrane. FASEB J 2012; 26:219–228 [View Article] [PubMed]
     [Google Scholar]
  20. Li GW, Burkhardt D, Gross C, Weissman JS. Quantifying absolute protein synthesis rates reveals principles underlying allocation of cellular resources. Cell 2014; 157:624–635 [View Article] [PubMed]
     [Google Scholar]
  21. Lazzaroni JC, Germon P, Ray MC, Vianney A. The Tol proteins of Escherichia coli and their involvement in the uptake of biomolecules and outer membrane stability. FEMS Microbiol Lett 1999; 177:191–197 [View Article] [PubMed]
     [Google Scholar]
  22. Cascales E, Bernadac A, Gavioli M, Lazzaroni JC, Lloubes R. Pal lipoprotein of Escherichia coli plays a major role in outer membrane integrity. J Bacteriol 2002; 184:754–759 [View Article] [PubMed]
     [Google Scholar]
  23. Bernadac A, Gavioli M, Lazzaroni JC, Raina S, Lloubès R. Escherichia coli Tol-Pal mutants form outer membrane vesicles. J Bacteriol 1998; 180:4872–4878 [View Article] [PubMed]
     [Google Scholar]
  24. Gerding MA, Ogata Y, Pecora ND, Niki H, de Boer PAJ. The trans-envelope Tol-Pal complex is part of the cell division machinery and required for proper outer-membrane invagination during cell constriction in E. coli. . Mol Microbiol 2007; 63:1008–1025 [View Article] [PubMed]
     [Google Scholar]
  25. Meury J, Devilliers G. Impairment of cell division in tolA mutants of Escherichia coli at low and high medium osmolarities. Biol Cell 1999; 91:67–75 [PubMed]
     [Google Scholar]
  26. Petiti M, Serrano B, Faure L, Lloubes R, Mignot T et al. Tol energy-driven localization of Pal and anchoring to the peptidoglycan promote outer-membrane constriction. J Mol Biol 2019; 431:3275–3288 [View Article] [PubMed]
     [Google Scholar]
  27. Baccelli P, Rachedi R, Serrano B, Petiti M, Bernard CS et al. Timing of TolA and TolQ recruitment at the septum depends on the functionality of the Tol-Pal system. J Mol Biol 2022; 434:167519 [View Article] [PubMed]
     [Google Scholar]
  28. Hale CA, Persons L, de Boer PAJ. Recruitment of the TolA protein to cell constriction sites in Escherichia coli via three separate mechanisms, and a critical role for FtsWI activity in recruitment of both TolA and TolQ. J Bacteriol 2022; 204:e00464-21 [View Article] [PubMed]
     [Google Scholar]
  29. Wojdyla JA, Cutts E, Kaminska R, Papadakos G, Hopper JTS et al. Structure and function of the Escherichia coli Tol-Pal stator protein TolR. J Biol Chem 2015; 290:26675–26687 [View Article] [PubMed]
     [Google Scholar]
  30. Rassam P, Long KR, Kaminska R, Williams DJ, Papadakos G et al. Intermembrane crosstalk drives inner-membrane protein organization in Escherichia coli. Nat Commun 2018; 9:1082 [View Article] [PubMed]
     [Google Scholar]
  31. Datsenko KA, Wanner BL. One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci U S A 2000; 97:6640–6645 [View Article] [PubMed]
     [Google Scholar]
  32. Banzhaf M, Yau HC, Verheul J, Lodge A, Kritikos G et al. Outer membrane lipoprotein NlpI scaffolds peptidoglycan hydrolases within multi-enzyme complexes in Escherichia coli. EMBO J 2020; 39:e102246 [View Article] [PubMed]
     [Google Scholar]
  33. Adams JN, Luria SE. Transduction by bacteriophage P1: abnormal phage function of the transducing particles. Proc Natl Acad Sci U S A 1958; 44:590–594 [View Article] [PubMed]
     [Google Scholar]
  34. Mori H, Baba T, Yokoyama K, Takeuchi R, Nomura W et al. Identification of essential genes and synthetic lethal gene combinations in Escherichia coli K-12. In Lu LJ. ed Gene Essentiality New York, NY: Springer New York; 2015 pp 45–65 [View Article]
     [Google Scholar]
  35. Vischer NOE, Verheul J, Postma M, van den Berg van Saparoea B, Galli E et al. Cell age dependent concentration of Escherichia coli divisome proteins analyzed with ImageJ and ObjectJ. Front Microbiol 2015; 6:586 [View Article] [PubMed]
     [Google Scholar]
  36. Buddelmeijer N, Aarsman M, den Blaauwen T. Immunolabeling of proteins in situ in Escherichia coli K12 strains. Bio Protoc 2013; 3: [View Article]
     [Google Scholar]
  37. Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y et al. Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol Syst Biol 2006; 2: [View Article] [PubMed]
     [Google Scholar]
  38. Hocking J, Priyadarshini R, Takacs CN, Costa T, Dye NA et al. Osmolality-dependent relocation of penicillin-binding protein PBP2 to the division site in Caulobacter crescentus. J Bacteriol 2012; 194:3116–3127 [View Article] [PubMed]
     [Google Scholar]
  39. van der Ploeg R, Verheul J, Vischer NOE, Alexeeva S, Hoogendoorn E et al. Colocalization and interaction between elongasome and divisome during a preparative cell division phase in Escherichia coli: preparing cell division. Mol Microbiol 2013; 87:1074–1087 [View Article] [PubMed]
     [Google Scholar]
  40. Gray AN, Egan AJ, Veer IL, Verheul J, Colavin A et al. Coordination of peptidoglycan synthesis and outer membrane constriction during Escherichia coli cell division. eLife 2015; 4:e07118
     [Google Scholar]
  41. Den Blaauwen T, Buddelmeijer N, Aarsman MEG, Hameete CM, Nanninga N. Timing of FtsZ assembly in Escherichia coli. J Bacteriol 1999; 181:5167–5175 [View Article] [PubMed]
     [Google Scholar]
  42. Jeon WJ, Cho H. A cell wall hydrolase meph is negatively regulated by proteolysis involving Prc and Nlpi in Escherichia coli. Front Microbiol 2022; 13:878049
     [Google Scholar]
  43. Bertsche U, Kast T, Wolf B, Fraipont C, Aarsman MEG et al. Interaction between two murein (peptidoglycan) synthases, PBP3 and PBP1B, in Escherichia coli. Mol Microbiol 2006; 61:675–690 [View Article] [PubMed]
     [Google Scholar]
  44. Verheul J, Lodge A, Yau HCL, Liu X, Boelter G et al. Early midcell localization of Escherichia coli PBP4 supports the function of peptidoglycan amidases. PLoS Genet 2022; 18:e1010222 [View Article] [PubMed]
     [Google Scholar]
  45. Aarsman MEG, Piette A, Fraipont C, Vinkenvleugel TMF, Nguyen-Distèche M et al. Maturation of the Escherichia coli divisome occurs in two steps: Maturation of the divisome. Mol Microbiol 2005; 55:1631–1645 [View Article] [PubMed]
     [Google Scholar]
  46. Goehring NW, Beckwith J. Diverse paths to midcell: assembly of the bacterial cell division machinery. Curr Biol 2005; 15:R514–R526 [View Article] [PubMed]
     [Google Scholar]
  47. den Blaauwen T, Hamoen LW, Levin PA. The divisome at 25: the road ahead. Curr Opin Microbiol 2017; 36:85–94 [View Article]
     [Google Scholar]
  48. Gogol EB, Rhodius VA, Papenfort K, Vogel J, Gross CA. Small RNAs endow a transcriptional activator with essential repressor functions for single-tier control of a global stress regulon. Proc Natl Acad Sci U S A 2011; 108:12875–12880 [View Article] [PubMed]
     [Google Scholar]
  49. Melamed S, Peer A, Faigenbaum-Romm R, Gatt YE, Reiss N et al. Global mapping of small RNA-target interactions in bacteria. Mol Cell 2016; 63:884–897 [View Article] [PubMed]
     [Google Scholar]
  50. Kaul M, Meher SK, Reddy M. Glycan strand cleavage by a lytic transglycosylase, MltD contributes to the expansion of peptidoglycan in Escherichia coli. BioRxiv Microbiol 2023 [View Article]
     [Google Scholar]
  51. Taschner PE, Huls PG, Pas E, Woldringh CL. Division behavior and shape changes in isogenic ftsZ, ftsQ, ftsA, pbpB, and ftsE cell division mutants of Escherichia coli during temperature shift experiments. J Bacteriol 1988; 170:1533–1540 [View Article] [PubMed]
     [Google Scholar]
  52. Mohammadi T, Ploeger GEJ, Verheul J, Comvalius AD, Martos A et al. The GTPase activity of Escherichia coli FtsZ determines the magnitude of the FtsZ polymer bundling by ZapA in vitro. . Biochemistry 2009; 48:11056–11066 [View Article] [PubMed]
     [Google Scholar]
  53. Koppelman C, Aarsman MEG, Postmus J, Pas E, Muijsers AO et al. R174 of Escherichia coli FtsZ is involved in membrane interaction and protofilament bundling, and is essential for cell division. BioRxiv Microbiol 2004; 51:645–657 [View Article]
     [Google Scholar]
/content/journal/acmi/10.1099/acmi.0.000759.v3
Loading
Cell division cycle fluctuation of Pal concentration in Escherichia coli
Access Microbiology 6, 000759.v3 (2024); https://doi.org/10.1099/acmi.0.000759.v3
/content/journal/acmi/10.1099/acmi.0.000759.v3
/content/journal/acmi/10.1099/acmi.0.000759.v3
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error