Survives with a Minimal Peptidoglycan Synthesis Machine but Sacrifices Virulence and Antibiotic Resistance

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Peptidoglycan forms the stress-bearing sacculus that prevents lysis of bacteria due to turgor pressure. The integrity of peptidoglycan is therefore essential for bacterial survival and its synthesis is the target of many important antibiotics, such as penicillin. The final steps of peptidoglycan synthesis are catalyzed by penicillin-binding proteins, enzymes that are proposed to work in multi-enzyme complexes. We show that seven of the nine genes encoding peptidoglycan synthesis enzymes can be deleted from the Staphylococcus aureus genome without affecting normal growth and cell morphology in vitro, identifying the minimal peptidoglycan synthesis machinery of this organism. Identification of minimal machineries is key for synthetic biology efforts towards the design of systems with reduced complexity. However, the non-essential peptidoglycan synthetic proteins are important for survival of S. aureus in more challenging environments, such as in the presence of antibiotics that target cell wall synthesis or within the host, as shown by the inability of the mutant strain to establish a successful infection and kill Drosophila flies.

Vyšlo v časopise: Survives with a Minimal Peptidoglycan Synthesis Machine but Sacrifices Virulence and Antibiotic Resistance. PLoS Pathog 11(5): e32767. doi:10.1371/journal.ppat.1004891
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1004891

Souhrn

Zdroje

1. Holtje JV (1996) A hypothetical holoenzyme involved in the replication of the murein sacculus of Escherichia coli. Microbiology 142: 1911–1918. 8760905

2. Holtje JV (1998) Growth of the stress-bearing and shape-maintaining murein sacculus of Escherichia coli. Microbiology and Molecular Biology Reviews 62: 181–203. 9529891

3. Scheffers DJ, Pinho MG (2005) Bacterial cell wall synthesis: New insights from localization studies. Microbiology and Molecular Biology Reviews 69: 585–607. 16339737

4. Hartman BJ, Tomasz A (1984) Low-Affinity Penicillin-Binding Protein Associated with Beta-Lactam Resistance in Staphylococcus aureus. Journal of Bacteriology 158: 513–516. 6563036

5. Pereira SFF, Henriques AO, Pinho MG, de Lencastre H, Tomasz A (2007) Role of PBP1 in cell division of Staphylococcus aureus. Journal of Bacteriology 189: 3525–3531. 17307860

6. Pereira SFF, Henriques AO, Pinho MG, de Lencastre H, Tomasz A (2009) Evidence for a dual role of PBP1 in the cell division and cell separation of Staphylococcus aureus. Molecular Microbiology 72: 895–904. doi: 10.1111/j.1365-2958.2009.06687.x 19400776

7. Pinho MG, Filipe SR, De Lencastre H, Tomasz A (2001) Complementation of the essential peptidoglycan transpeptidase function of penicillin-binding protein 2 (PBP2) by the drug resistance protein PBP2A in Staphylococcus aureus. Journal of Bacteriology 183: 6525–6531. 11673420

8. Pinho MG, de Lencastre H, Tomasz A (2001) An acquired and a native penicillin-binding protein cooperate in building the cell wall of drug-resistant staphylococci. Proceedings of the National Academy of Sciences of the United States of America 98: 10886–10891. 11517340

9. Pinho MG, de Lencastre H, Tomasz A (2000) Cloning, characterization, and inactivation of the gene pbpC, encoding penicillin-binding protein 3 of Staphylococcus aureus. Journal of Bacteriology 182: 1074–1079. 10648534

10. Curtis NAC, Hayes MV, Wyke AW, Ward JB (1980) A mutant of Staphylococcus aureus H lacking penicillin-binding protein 4 and transpeptidase activity in vitro. Fems Microbiology Letters 9: 263–266.

11. Memmi G, Filipe SR, Pinho MG, Fu ZB, Cheung A (2008) Staphylococcus aureus PBP4 is essential for beta-lactam resistance in community-acquired methicillin-resistant strains. Antimicrobial Agents and Chemotherapy 52: 3955–3966. doi: 10.1128/AAC.00049-08 18725435

12. Reed P, Veiga H, Jorge AM, Terrak M, Pinho MG (2011) Monofunctional transglycosylases are not essential for Staphylococcus aureus cell wall synthesis. Journal of Bacteriology 193: 2549–2556. doi: 10.1128/JB.01474-10 21441517

13. Coutinho PM, Deleury E, Davies GJ, Henrissat B (2003) An evolving hierarchical family classification for glycosyltransferases. Journal of Molecular Biology 328: 307–317. 12691742

14. Kuroda M, Ohta T, Uchiyama I, Baba T, Yuzawa H, et al. (2001) Whole genome sequencing of meticillin-resistant Staphylococcus aureus. Lancet 357: 1225–1240. 11418146

15. Komatsuzawa H, Ohta K, Sugai M, Fujiwara T, Glanzmann P, et al. (2000) Tn551-mediated insertional inactivation of the fmtB gene encoding a cell wall-associated protein abolishes methicillin resistance in Staphylococcus aureus. Journal of Antimicrobial Chemotherapy 45: 421–431. 10896508

16. Fan X, Liu YH, Smith D, Konermann L, Siu KWM, et al. (2007) Diversity of penicillin-binding proteins—Resistance factor FmtA of Staphylococcus aureus. Journal of Biological Chemistry 282: 35143–35152. 17925392

17. Komatsuzawa H, Ohta K, Labischinski H, Sugai M, Suginaka H (1999) Characterization of fmtA, a gene that modulates the expression of methicillin resistance in Staphylococcus aureus. Antimicrobial Agents and Chemotherapy 43: 2121–2125. 10471551

18. Boucher HW, Corey GR (2008) Epidemiology of methicillin-resistant Staphylococcus aureus. Clinical Infectious Diseases 46: S344–S349. doi: 10.1086/533590 18462089

19. Eddy SR (1998) Profile hidden Markov models. Bioinformatics 14: 755–763. 9918945

20. Boneca IG, de Reuse H, Epinat JC, Pupin M, Labigne A, et al. (2003) A revised annotation and comparative analysis of Helicobacter pylori genomes. Nucleic Acids Research 31: 1704–1714. 12626712

21. El Ghachi M, Mattei PJ, Ecobichon C, Martins A, Hoos S, et al. (2011) Characterization of the elongasome core PBP2: MreC complex of Helicobacter pylori. Molecular Microbiology 82: 68–86. doi: 10.1111/j.1365-2958.2011.07791.x 21801243

22. Boneca IG, Ecobichon C, Chaput C, Mathieu A, Guadagnini S, et al. (2008) Development of inducible systems to engineer conditional mutants of essential genes of Helicobacter pylori. Applied Environmental Microbiology 74: 2095–2102. doi: 10.1128/AEM.01348-07 18245237

23. Matthews P, Tomasz A (1990) Insertional inactivation of the mec gene in a transposon mutant of a methicillin-resistant clinical isolate of Staphylococcus aureus. Antimicrobial Agents and Chemotherapy 34: 1777–1779. 2178337

24. Pereira AR, Reed P, Veiga H, Pinho MG (2013) The Holliday junction resolvase RecU is required for chromosome segregation and DNA damage repair in Staphylococcus aureus. BMC Microbiology 13: 18. doi: 10.1186/1471-2180-13-18 23356868

25. Tam R, Saier MH Jr. (1993) Structural, functional, and evolutionary relationships among extracellular solute-binding receptors of bacteria. Microbiology Reviews 57: 320–346. 8336670

26. Kuroda M, Kuroda H, Oshima T, Takeuchi F, Mori H, et al. (2003) Two-component system VraSR positively modulates the regulation of cell-wall biosynthesis pathway in Staphylococcus aureus. Molecular Microbiology 49: 807–821. 12864861

27. Pinho MG, Errington J (2005) Recruitment of penicillin-binding protein PBP2 to the division site of Staphylococcus aureus is dependent on its transpeptidation substrates. Molecular Microbiology 55: 799–807. 15661005

28. Kuru E, Hughes HV, Brown PJ, Hall E, Tekkam S, et al. (2012) In situ probing of newly synthesized peptidoglycan in live bacteria with fluorescent D-amino acids. Angewandte Chemie-International Edition 51: 12519–12523. doi: 10.1002/anie.201206749 23055266

29. Pinho MG, Errington J (2003) Dispersed mode of Staphylococcus aureus cell wall synthesis in the absence of the division machinery. Molecular Microbiology 50: 871–881. 14617148

30. Steele VR, Bottomley AL, Garcia-Lara J, Kasturiarachchi J, Foster SJ (2011) Multiple essential roles for EzrA in cell division of Staphylococcus aureus. Molecular Microbiology 80: 542–555. doi: 10.1111/j.1365-2958.2011.07591.x 21401734

31. Wyke AW, Ward JB, Hayes MV, Curtis NAC (1981) A Role in vivo for Penicillin-Binding Protein 4 of Staphylococcus aureus. European Journal of Biochemistry 119: 389–393. 7308191

32. Boneca IG, Huang ZH, Gage DA, Tomasz A (2000) Characterization of Staphylococcus aureus cell wall glycan strands, evidence for a new beta-N-acetylglucosaminidase activity. Journal of Biological Chemistry 275: 9910–9918. 10744664

33. Atilano ML, Yates J, Glittenberg M, Filipe SR, Ligoxygakis P (2011) Wall teichoic acids of Staphylococcus aureus limit recognition by the Drosophila peptidoglycan recognition protein-SA to promote pathogenicity. Plos Pathogens 7:E1002421. doi: 10.1371/journal.ppat.1002421 22144903

34. Kounatidis I, Ligoxygakis P (2012) Drosophila as a model system to unravel the layers of innate immunity to infection. Open Biology 2: 120075. doi: 10.1098/rsob.120075 22724070

35. Chang CI, Pili-Floury S, Herve M, Parquet C, Chelliah Y, et al. (2004) A Drosophila pattern recognition receptor contains a peptidoglycan docking groove and unusual L,D-carboxypeptidase activity. PLoS Biology 2: E277. 15361936

36. Bera A, Biswas R, Herbert S, Kulauzovic E, Weidenmaier C, et al. (2007) Influence of wall teichoic acid on lysozyme resistance in Staphylococcus aureus. Journal of Bacteriology 189: 280–283. 17085565

37. Atilano ML, Pereira PM, Yates J, Reed P, Veiga H, et al. (2010) Teichoic acids are temporal and spatial regulators of peptidoglycan cross-linking in Staphylococcus aureus. Proceedings of the National Academy of Sciences of the United States of America 107: 18991–18996. doi: 10.1073/pnas.1004304107 20944066

38. Forsyth RA, Haselbeck RJ, Ohlsen KL, Yamamoto RT, Xu H, et al. (2002) A genome-wide strategy for the identification of essential genes in Staphylococcus aureus. Molecular Microbiology 43: 1387–1400. 11952893

39. Labischinski H, Goodell EW, Goodell A, Hochberg ML (1991) Direct proof of a more-than-single-layered peptidoglycan architecture of Escherichia coli W7: a neutron small-angle scattering study. Journal of Bacteriology 173: 751–756. 1987162

40. Typas A, Banzhaf M, Gross CA, Vollmer W (2012) From the regulation of peptidoglycan synthesis to bacterial growth and morphology. Nature Reviews Microbiology 10: 123–136. doi: 10.1038/nrmicro2677 22203377

41. Kelman Z, Odonnell M (1995) DNA-Polymerase-III Holoenzyme—Structure and function of a chromosomal replicating machine. Annual Review of Biochemistry 64: 171–200. 7574479

42. Borukhov S, Nudler E (2003) RNA polymerase holoenzyme: structure, function and biological implications. Current Opinion in Microbiology 6: 93–100. 12732296

43. Giesbrecht P, Kersten T, Maidhof H, Wecke J (1998) Staphylococcal cell wall: Morphogenesis and fatal variations in the presence of penicillin. Microbiology and Molecular Biology Reviews 62: 1371–1414. 9841676

44. Goehring NW, Beckwith J (2005) Diverse paths to midcell: Assembly of the bacterial cell division machinery. Current Biology 15: R514–R526. 16005287

45. Vicente M, Rico AI (2006) The order of the ring: assembly of Escherichia coli cell division components. Molecular Microbiology 61: 5–8. 16824090

46. McPherson DC, Popham DL (2003) Peptidoglycan synthesis in the absence of class A penicillin-binding proteins in Bacillus subtilis. Journal of Bacteriology 185: 1423–1431. 12562814

47. Rice LB, Carias LL, Rudin S, Hutton R, Marshall S, et al. (2009) Role of class A penicillin-binding proteins in the expression of beta-lactam resistance in Enterococcus faecium. Journal of Bacteriology 191: 3649–3656. doi: 10.1128/JB.01834-08 19304851

48. Antignac A, Sieradzki K, Tomasz A (2007) Perturbation of cell wall synthesis suppresses autolysis in Staphylococcus aureus: evidence for coregulation of cell wall synthetic and hydrolytic enzymes. Journal of Bacteriology 189: 7573–7580. 17827298

49. Strobel W, Moll A, Kiekebusch D, Klein KE, Thanbichler M (2014) Function and localization dynamics of bifunctional penicillin-binding proteins in Caulobacter crescentus. Journal of Bacteriology 196: 1627–1639. doi: 10.1128/JB.01194-13 24532768

50. Sakoulas G, Okumura CY, Thienphrapa W, Olson J, Nonejuie P, et al. (2014) Nafcillin enhances innate immune-mediated killing of methicillin-resistant Staphylococcus aureus. Journal of Molecular Medicine (Berlin) 92: 139–149. doi: 10.1007/s00109-013-1100-7 24297496

51. Gough J, Karplus K, Hughey R, Chothia C (2001) Assignment of homology to genome sequences using a library of hidden Markov models that represent all proteins of known structure. Journal of Molecular Biology 313: 903–919. 11697912

52. Wu M, Scott AJ (2012) Phylogenomic analysis of bacterial and archaeal sequences with AMPHORA2. Bioinformatics 28: 1033–1034. doi: 10.1093/bioinformatics/bts079 22332237

53. Guindon S, Gascuel O (2003) A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Systematic Biology 52: 696–704. 14530136

54. Wu DY, Hugenholtz P, Mavromatis K, Pukall R, Dalin E, et al. (2009) A phylogeny-driven genomic encyclopaedia of Bacteria and Archaea. Nature 462: 1056–1060. doi: 10.1038/nature08656 20033048

55. Langmead B, Trapnell C, Pop M, Salzberg SL (2009) Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome Biology 10: R25. doi: 10.1186/gb-2009-10-3-r25 19261174

56. Filipe SR, Tomasz A, Ligoxygakis P (2005) Requirements of peptidoglycan structure that allow detection by the Drosophila Toll pathway. Embo Reports 6: 327–333. 15791270

57. Jorge AM, Hoiczyk E, Gomes JP, Pinho MG (2011) EzrA contributes to the regulation of cell size in Staphylococcus aureus. Plos One 6:E27542. doi: 10.1371/journal.pone.0027542 22110668

58. Michel T, Reichhart JM, Hoffmann JA, Royet J (2001) Drosophila Toll is activated by Gram-positive bacteria through a circulating peptidoglycan recognition protein. Nature 414: 756–759. 11742401

59. Mackay TF, Richards S, Stone EA, Barbadilla A, Ayroles JF, et al. (2012) The Drosophila melanogaster genetic reference panel. Nature 482: 173–178. doi: 10.1038/nature10811 22318601