Protein Complexes and Proteolytic Activation of the Cell Wall Hydrolase RipA Regulate Septal Resolution in Mycobacteria

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Peptidoglycan hydrolases are a double-edged sword. They are required for normal cell division, but when dysregulated can become autolysins lethal to bacteria. How bacteria ensure that peptidoglycan hydrolases function only in the correct spatial and temporal context remains largely unknown. Here, we demonstrate that dysregulation converts the essential mycobacterial peptidoglycan hydrolase RipA to an autolysin that compromises cellular structural integrity. We find that mycobacteria control RipA activity through two interconnected levels of regulation in vivo—protein interactions coordinate PG hydrolysis, while proteolysis is necessary for RipA enzymatic activity. Dysregulation of RipA protein complexes by treatment with a peptidoglycan synthase inhibitor leads to excessive RipA activity and impairment of correct morphology. Furthermore, expression of a RipA dominant negative mutant or of differentially processed RipA homologues reveals that RipA is produced as a zymogen, requiring proteolytic processing for activity. The amount of RipA processing differs between fast-growing and slow-growing mycobacteria and correlates with the requirement for peptidoglycan hydrolase activity in these species. Together, the complex picture of RipA regulation is a part of a growing paradigm for careful control of cell wall hydrolysis by bacteria during growth, and may represent a novel target for chemotherapy development.

Vyšlo v časopise: Protein Complexes and Proteolytic Activation of the Cell Wall Hydrolase RipA Regulate Septal Resolution in Mycobacteria. PLoS Pathog 9(2): e32767. doi:10.1371/journal.ppat.1003197
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1003197

Souhrn

Zdroje

1. WHO (2010) Global Tuberculosis Control 2010. Geneva: WHO Press.

2. UdwadiaZF, AmaleRA, AjbaniKK, RodriguesC (2012) Totally drug-resistant tuberculosis in India. Clin Infect Dis 54 : 579–581.

3. SauvageE, KerffF, TerrakM, AyalaJA, CharlierP (2008) The penicillin-binding proteins: structure and role in peptidoglycan biosynthesis. FEMS Microbiol Rev 32 : 234–258.

4. VollmerW, JorisB, CharlierP, FosterS (2008) Bacterial peptidoglycan (murein) hydrolases. FEMS Microbiol Rev 32 : 259–286.

5. MeiselU, HoltjeJ-V, VollmerW (2003) Overproduction of Inactive Variants of the Murein Synthase PBP1B Causes Lysis in Escherichia coli. J Bacteriol 185 : 5342–5348.

6. LegareeBA, AdamsCB, ClarkeAJ (2007) Overproduction of Penicillin-Binding Protein 2 and Its Inactive Variants Causes Morphological Changes and Lysis in Escherichia coli. J Bacteriol 189 : 4975–4983.

7. HeidrichC, UrsinusA, BergerJ, SchwarzH, HoltjeJ-V (2002) Effects of Multiple Deletions of Murein Hydrolases on Viability, Septum Cleavage, and Sensitivity to Large Toxic Molecules in Escherichia coli. J Bacteriol 184 : 6093–6099.

8. den BlaauwenT, de PedroMA, Nguyen-DistècheM, AyalaJA (2008) Morphogenesis of rod-shaped sacculi. FEMS Microbiol Rev 32 : 321–344.

9. UeharaT, ParzychKR, DinhT, BernhardtTG (2010) Daughter cell separation is controlled by cytokinetic ring-activated cell wall hydrolysis. EMBO J 29 : 1412–1422.

10. Paradis-BleauC, MarkovskiM, UeharaT, LupoliTJ, WalkerS, et al. (2010) Lipoprotein cofactors located in the outer membrane activate bacterial cell wall polymerases. Cell 143 : 1110–1120.

11. VollmerW, von RechenbergM, HöltjeJ-V (1999) Demonstration of Molecular Interactions between the Murein Polymerase PBP1B, the Lytic Transglycosylase MltA, and the Scaffolding Protein MipA of Escherichia coli. J Biol Chem 274 : 6726–6734.

12. RomeisT, HöltjeJV (1994) Specific interaction of penicillin-binding proteins 3 and 7/8 with soluble lytic transglycosylase in Escherichia coli. J Biol Chem 269 : 21603–21607.

13. BertscheU, KastT, WolfB, FraipontC, AarsmanME, et al. (2006) Interaction between two murein (peptidoglycan) synthases, PBP3 and PBP1B, in Escherichia coli. Mol Microbiol 61 : 675–690.

14. BöthD, SchneiderG, SchnellR (2011) Peptidoglycan remodeling in Mycobacterium tuberculosis: comparison of structures and catalytic activities of RipA and RipB. J Mol Biol 413 : 247–260.

15. PilgrimS, Kolb-MaurerA, GentschevI, GoebelW, KuhnM (2003) Deletion of the Gene Encoding p60 in Listeria monocytogenes Leads to Abnormal Cell Division and Loss of Actin-Based Motility. Infect Immun 71 : 3473–3484.

16. GaoL-Y, PakM, KishR, KajiharaK, BrownEJ (2006) A mycobacterial operon essential for virulence in vivo and invasion and intracellular persistence in macrophages. Infect Immun 74 : 1757–1767.

17. SassettiCM, BoydDH, RubinEJ (2003) Genes required for mycobacterial growth defined by high density mutagenesis. Molecular Microbiology 48 : 77–84.

18. HettEC, ChaoMC, DengLL, RubinEJ (2008) A mycobacterial enzyme essential for cell division synergizes with resuscitation-promoting factor. PLoS Pathog 4: e1000001.

19. HeidrichC, TemplinMF, UrsinusA, MerdanovicM, BergerJ, 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.

20. HugonnetJ-E, TremblayLW, BoshoffHI, BarryCE, BlanchardJS (2009) Meropenem-Clavulanate Is Effective Against Extensively Drug-Resistant Mycobacterium tuberculosis. Science 323 : 1215–1218.

21. GuptaR, LavollayM, MainardiJ-L, ArthurM, BishaiWR, et al. (2010) The Mycobacterium tuberculosis protein LdtMt2 is a nonclassical transpeptidase required for virulence and resistance to amoxicillin. Nat Med 16 : 466–469.

22. SumitaY, FakasawaM (1995) Potent activity of meropenem against Escherichia coli arising from its simultaneous binding to penicillin-binding proteins 2 and 3. Journal of Antimicrobial Chemotherapy 36 : 53–64.

23. Billman-JacobeH, HaitesRE, CoppelRL (1999) Characterization of a Mycobacterium smegmatis Mutant Lacking Penicillin Binding Protein 1. Antimicrob Agents Chemother 43 : 3011–3013.

24. HettEC, ChaoMC, RubinEJ (2010) Interaction and modulation of two antagonistic cell wall enzymes of mycobacteria. PLoS Pathog 6: e1001020.

25. RuggieroA, MarascoD, SquegliaF, SoldiniS, PedoneE, et al. (2010) Structure and Functional Regulation of RipA, a Mycobacterial Enzyme Essential for Daughter Cell Separation. Structure 18 : 1184–1190.

26. SlaydenRA, BelisleJT (2009) Morphological features and signature gene response elicited by inactivation of FtsI in Mycobacterium tuberculosis. Journal of Antimicrobial Chemotherapy 63 : 451–457.

27. HettEC, ChaoMC, SteynAJ, FortuneSM, DengLL, et al. (2007) A partner for the resuscitation-promoting factors of Mycobacterium tuberculosis. Molecular Microbiology 66 : 658–668.

28. ZhangYJ, IoergerTR, HuttenhowerC, LongJE, SassettiCM, et al. (2012) Global assessment of genomic regions required for growth in Mycobacterium tuberculosis. PLoS Pathog 8: e1002946.

29. BreciLA, TabbDL, YatesJR3rd, WysockiVH (2003) Cleavage N-terminal to proline: analysis of a database of peptide tandem mass spectra. Anal Chem 75 : 1963–1971.

30. BublitzM, PolleL, HollandC, HeinzDW, NimtzM, et al. (2009) Structural basis for autoinhibition and activation of Auto, a virulence-associated peptidoglycan hydrolase of Listeria monocytogenes. Molecular Microbiology 71 : 1509–1522.

31. ReddyTB, RileyR, WymoreF, MontgomeryP, DeCaprioD, et al. (2009) TB database: an integrated platform for tuberculosis research. Nucleic Acids Res 37: D499–508.

32. PlocinskaR, PurushothamG, SarvaK, VadrevuIS, PandeetiEV, et al. (2012) Septal localization of the Mycobacterium tuberculosis MtrB sensor kinase promotes MtrA regulon expression. J Biol Chem 287 : 23887–23899.

33. SegevE, SmithY, Ben-YehudaS (2012) RNA dynamics in aging bacterial spores. Cell 148 : 139–149.

34. VoskuilMI, ViscontiKC, SchoolnikGK (2004) Mycobacterium tuberculosis gene expression during adaptation to stationary phase and low-oxygen dormancy. Tuberculosis (Edinb) 84 : 218–227.

35. ShahIM, DworkinJ (2010) Induction and regulation of a secreted peptidoglycan hydrolase by a membrane Ser/Thr kinase that detects muropeptides. Molecular Microbiology 75 : 1232–1243.

36. ShahIM, LaaberkiM-H, PophamDL, DworkinJ (2008) A Eukaryotic-like Ser/Thr Kinase Signals Bacteria to Exit Dormancy in Response to Peptidoglycan Fragments. Cell 135 : 486–496.

37. DowningKJ, MischenkoVV, ShleevaMO, YoungDI, YoungM, et al. (2005) Mutants of Mycobacterium tuberculosis lacking three of the five rpf-like genes are defective for growth in vivo and for resuscitation in vitro. Infection and Immunity 73 : 3038–3043.

38. BiketovS, PotapovV, GaninaE, DowningK, KanaB, et al. (2007) The role of resuscitation promoting factors in pathogenesis and reactivation of Mycobacterium tuberculosis during intra-peritoneal infection in mice. BMC Infectious Diseases 7 : 146–153.

39. MirM, AsongJ, LiX, CardotJ, BoonsGJ, et al. (2011) The extracytoplasmic domain of the Mycobacterium tuberculosis Ser/Thr kinase PknB binds specific muropeptides and is required for PknB localization. PLoS Pathog 7: e1002182.

40. EhrtS, GuoXV, HickeyCM, RyouM, MonteleoneM, et al. (2005) Controlling gene expression in mycobacteria with anhydrotetracycline and Tet repressor. Nucleic Acids Res 33: e21.