1887

Abstract

Bacterial cell division is mediated by a protein complex known as the divisome. Many protein–protein interactions in the divisome have been characterized. In this report, we analyse the role of the PASTA (Penicillin-binding protein And Serine Threonine kinase Associated) domains of PBP2B. PBP2B itself is essential and cannot be deleted, but removing the PBP2B PASTA domains results in impaired cell division and a heat-sensitive phenotype. This resembles the deletion of , a known interaction partner of PBP2B. Bacterial two-hybrid and co-immunoprecipitation analyses show that the interaction between PBP2B and DivIB is weakened when the PBP2B PASTA domains are removed. Combined, our results show that the PBP2B PASTA domains are required to strengthen the interaction between PBP2B and DivIB.

Funding
This study was supported by the:
  • Dirk-Jan Scheffers , Nederlandse Organisatie voor Wetenschappelijk Onderzoek , (Award 864.09.010)
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2020-08-04
2020-10-29
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References

  1. Monteiro JM, Pereira AR, Reichmann NT, Saraiva BM, Fernandes PB et al. Peptidoglycan synthesis drives an FtsZ-treadmilling-independent step of cytokinesis. Nature 2018; 554:528–532 [CrossRef][PubMed]
    [Google Scholar]
  2. den Blaauwen T, Hamoen LW, Levin PA. The divisome at 25: the road ahead. Curr Opin Microbiol 2017; 36:85–94 [CrossRef][PubMed]
    [Google Scholar]
  3. Taguchi A, Welsh MA, Marmont LS, Lee W, Sjodt M et al. FtsW is a peptidoglycan polymerase that is functional only in complex with its cognate penicillin-binding protein. Nat Microbiol 2019; 4:587–594 [CrossRef][PubMed]
    [Google Scholar]
  4. Zhao H, Patel V, Helmann JD, Dörr T. Don't let sleeping dogmas lie: new views of peptidoglycan synthesis and its regulation. Mol Microbiol 2017; 106:847–860 [CrossRef][PubMed]
    [Google Scholar]
  5. Sjodt M, Rohs PDA, Gilman MSA, Erlandson SC, Zheng S et al. Structural coordination of polymerization and crosslinking by a SEDS-bPBP peptidoglycan synthase complex. Nat Microbiol 2020; 5:813–820 [CrossRef][PubMed]
    [Google Scholar]
  6. Kobayashi K, Ehrlich SD, Albertini A, Amati G, Andersen KK et al. Essential Bacillus subtilis genes. Proc Natl Acad Sci USA 2003; 100:4678–4683 [CrossRef][PubMed]
    [Google Scholar]
  7. Daniel RA, Williams AM, Errington J. A complex four-gene operon containing essential cell division gene pbpB in Bacillus subtilis . J Bacteriol 1996; 178:2343–2350 [CrossRef][PubMed]
    [Google Scholar]
  8. Morales Angeles D, Liu Y, Hartman AM, Borisova M, de Sousa Borges A et al. Pentapeptide-rich peptidoglycan at the Bacillus subtilis cell-division site. Mol Microbiol 2017; 104:319333 [CrossRef][PubMed]
    [Google Scholar]
  9. Sassine J, Xu M, Sidiq KR, Emmins R, Errington J et al. Functional redundancy of division specific penicillin-binding proteins in Bacillus subtilis . Mol Microbiol 2017; 106:304–318 [CrossRef][PubMed]
    [Google Scholar]
  10. Peters K, Schweizer I, Beilharz K, Stahlmann C, Veening J-W et al. Streptococcus pneumoniae PBP2x mid-cell localization requires the C-terminal PASTA domains and is essential for cell shape maintenance. Mol Microbiol 2014; 92:733–755 [CrossRef][PubMed]
    [Google Scholar]
  11. Yeats C, Finn RD, Bateman A. The PASTA domain: a β-lactam-binding domain. Trends Biochem Sci 2002; 27:438440 [CrossRef][PubMed]
    [Google Scholar]
  12. Yanouri A, Daniel RA, Errington J, Buchanan CE. Cloning and sequencing of the cell division gene pbpB, which encodes penicillin-binding protein 2B in Bacillus subtilis . J Bacteriol 1993; 175:7604–7616 [CrossRef][PubMed]
    [Google Scholar]
  13. Maurer P, Todorova K, Sauerbier J, Hakenbeck R. Mutations in Streptococcus pneumoniae penicillin-binding protein 2x: importance of the C-terminal penicillin-binding protein and serine/threonine kinase-associated domains for beta-lactam binding. Microb Drug Resist 2012; 18:314–321 [CrossRef][PubMed]
    [Google Scholar]
  14. Bernardo-García N, Mahasenan KV, Batuecas MT, Lee M, Hesek D et al. Allostery, recognition of nascent peptidoglycan, and cross-linking of the cell wall by the essential penicillin-binding protein 2x of Streptococcus pneumoniae . ACS Chem Biol 2018; 13:694–702 [CrossRef][PubMed]
    [Google Scholar]
  15. Calvanese L, Falcigno L, Squeglia F, D'Auria G, Berisio R. Structural and dynamic features of PASTA domains with different functional roles. J Biomol Struct Dyn 2017; 35:2293–2300 [CrossRef][PubMed]
    [Google Scholar]
  16. Shah IM, Laaberki M-H, Popham DL, Dworkin J. A eukaryotic-like Ser/Thr kinase signals bacteria to exit dormancy in response to peptidoglycan fragments. Cell 2008; 135:486–496 [CrossRef][PubMed]
    [Google Scholar]
  17. Squeglia F, Marchetti R, Ruggiero A, Lanzetta R, Marasco D et al. Chemical basis of peptidoglycan discrimination by PrkC, a key kinase involved in bacterial resuscitation from dormancy. J Am Chem Soc 2011; 133:20676–20679 [CrossRef][PubMed]
    [Google Scholar]
  18. Ruggiero A, Squeglia F, Marasco D, Marchetti R, Molinaro A et al. X-ray structural studies of the entire extracellular region of the serine/threonine kinase PrkC from Staphylococcus aureus . Biochem J 2011; 435:33–41 [CrossRef][PubMed]
    [Google Scholar]
  19. Mir M, Asong J, Li X, Cardot J, Boons G-J et al. The extracytoplasmic domain of the Mycobacterium tuberculosis Ser/Thr kinase PknB binds specific muropeptides and is required for PknB localization. PLoS Pathog 2011; 7:e1002182 [CrossRef][PubMed]
    [Google Scholar]
  20. Barthe P, Mukamolova GV, Roumestand C, Cohen-Gonsaud M. The structure of PknB extracellular PASTA domain from Mycobacterium tuberculosis suggests a ligand-dependent kinase activation. Structure 2010; 18:606–615 [CrossRef][PubMed]
    [Google Scholar]
  21. Paracuellos P, Ballandras A, Robert X, Kahn R, Hervé M et al. The extended conformation of the 2.9-Å crystal structure of the three-PASTA domain of a Ser/Thr kinase from the human pathogen Staphylococcus aureus . J Mol Biol 2010; 404:847–858 [CrossRef][PubMed]
    [Google Scholar]
  22. Maestro B, Novaková L, Hesek D, Lee M, Leyva E et al. Recognition of peptidoglycan and β-lactam antibiotics by the extracellular domain of the Ser/Thr protein kinase StkP from Streptococcus pneumoniae . FEBS Lett 2011; 585:357–363 [CrossRef][PubMed]
    [Google Scholar]
  23. Morlot C, Bayle L, Jacq M, Fleurie A, Tourcier G et al. Interaction of penicillin-binding protein 2x and Ser/Thr protein kinase StkP, two key players in Streptococcus pneumoniae R6 morphogenesis. Mol Microbiol 2013; 90:88–102 [CrossRef][PubMed]
    [Google Scholar]
  24. Pensinger DA, Schaenzer AJ, Sauer J-D. Do shoot the messenger: PASTA kinases as virulence determinants and antibiotic targets. Trends Microbiol 2018; 26:56–69 [CrossRef][PubMed]
    [Google Scholar]
  25. Zucchini L, Mercy C, Garcia PS, Cluzel C, Gueguen-Chaignon V et al. PASTA repeats of the protein kinase StkP interconnect cell constriction and separation of Streptococcus pneumoniae . Nat Microbiol 2018; 3:197–209 [CrossRef][PubMed]
    [Google Scholar]
  26. Bond SR, Naus CC. RF-Cloning.org: an online tool for the design of restriction-free cloning projects. Nucleic Acids Res 2012; 40:W209–W213 [CrossRef][PubMed]
    [Google Scholar]
  27. Syvertsson S, Vischer NOE, Gao Y, Hamoen LW. When phase contrast fails: ChainTracer and NucTracer, two ImageJ methods for semi-automated single cell analysis using membrane or DNA staining. PLoS One 2016; 11:e0151267 [CrossRef][PubMed]
    [Google Scholar]
  28. Battesti A, Bouveret E. The bacterial two-hybrid system based on adenylate cyclase reconstitution in Escherichia coli . Methods 2012; 58:325–334 [CrossRef][PubMed]
    [Google Scholar]
  29. Scheffers D-J, Robichon C, Haan GJ, den Blaauwen T, Koningstein G et al. Contribution of the FtsQ transmembrane segment to localization to the cell division site. J Bacteriol 2007; 189:7273–7280 [CrossRef][PubMed]
    [Google Scholar]
  30. Zielińska A, Savietto A, de Sousa Borges A, Roelofsen JR, Hartman AM et al. Membrane fluidity controls peptidoglycan synthesis and MreB movement. bioRxiv 2019736819 [CrossRef]
    [Google Scholar]
  31. Daniel RA, Drake S, Buchanan CE, Scholle R, Errington J. The Bacillus subtilis spoVD gene encodes a mother-cell-specific penicillin-binding protein required for spore morphogenesis. J Mol Biol 1994; 235:209–220 [CrossRef]
    [Google Scholar]
  32. Libby EA, Goss LA, Dworkin J. The eukaryotic-like Ser/Thr kinase PrkC regulates the essential WalRK two-component system in Bacillus subtilis . PLoS Genet 2015; 11:e1005275 [CrossRef]
    [Google Scholar]
  33. Pompeo F, Foulquier E, Serrano B, Grangeasse C, Galinier A. Phosphorylation of the cell division protein GpsB regulates PrkC kinase activity through a negative feedback loop in Bacillus subtilis . Mol Microbiol 2015; 97:139–150 [CrossRef]
    [Google Scholar]
  34. Fay A, Meyer P, Dworkin J. Interactions between late-acting proteins required for peptidoglycan synthesis during sporulation. J Mol Biol 2010; 399:547–561 [CrossRef]
    [Google Scholar]
  35. Daniel RA, Errington J. Intrinsic instability of the essential cell division protein FtsL of Bacillus subtilis and a role for DivIB protein in FtsL turnover. Mol Microbiol 2000; 36:278–289 [CrossRef]
    [Google Scholar]
  36. Beall B, Lutkenhaus J. Nucleotide sequence and insertional inactivation of a Bacillus subtilis gene that affects cell division, sporulation, and temperature sensitivity. J Bacteriol 1989; 171:6821–6834 [CrossRef]
    [Google Scholar]
  37. Daniel RA, Noirot-Gros M-F, Noirot P, Errington J. Multiple interactions between the transmembrane division proteins of Bacillus subtilis and the role of FtsL instability in divisome assembly. J Bacteriol 2006; 188:7396–7404 [CrossRef]
    [Google Scholar]
  38. Rowland SL, Wadsworth KD, Robson SA, Robichon C, Beckwith J et al. Evidence from artificial septal targeting and site-directed mutagenesis that residues in the extracytoplasmic β domain of DivIB mediate its interaction with the divisomal transpeptidase PBP 2B. J Bacteriol 2010; 192:6116–6125 [CrossRef]
    [Google Scholar]
  39. Robichon C, King GF, Goehring NW, Beckwith J. Artificial septal targeting of Bacillus subtilis cell division proteins in Escherichia coli: an interspecies approach to the study of protein-protein interactions in multiprotein complexes. J Bacteriol 2008; 190:6048–6059 [CrossRef]
    [Google Scholar]
  40. Fenton AK, Manuse S, Flores-Kim J, Garcia PS, Mercy C et al. Phosphorylation-dependent activation of the cell wall synthase PBP2a in Streptococcus pneumoniae by MacP. Proc Natl Acad Sci USA 2018; 115:2812–2817 [CrossRef]
    [Google Scholar]
  41. Hamoen LW, Errington J. Polar targeting of DivIVA in Bacillus subtilis is not directly dependent on FtsZ or PBP 2B. J Bacteriol 2003; 185:693–697 [CrossRef]
    [Google Scholar]
  42. Karimova G, Pidoux J, Ullmann A, Ladant D. A bacterial two-hybrid system based on a reconstituted signal transduction pathway. Proc Natl Acad Sci USA 1998; 95:5752–5756 [CrossRef]
    [Google Scholar]
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