The Extracytoplasmic Domain of the Ser/Thr Kinase PknB Binds Specific Muropeptides and Is Required for PknB Localization

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The Mycobacterium tuberculosis Ser/Thr kinase PknB has been implicated in the regulation of cell growth and morphology in this organism. The extracytoplasmic domain of this membrane protein comprises four penicillin binding protein and Ser/Thr kinase associated (PASTA) domains, which are predicted to bind stem peptides of peptidoglycan. Using a comprehensive library of synthetic muropeptides, we demonstrate that the extracytoplasmic domain of PknB binds muropeptides in a manner dependent on the presence of specific amino acids at the second and third positions of the stem peptide, and on the presence of the sugar moiety N-acetylmuramic acid linked to the peptide. We further show that PknB localizes strongly to the mid-cell and also to the cell poles, and that the extracytoplasmic domain is required for PknB localization. In contrast to strong growth stimulation by conditioned medium, we observe no growth stimulation of M. tuberculosis by a synthetic muropeptide with high affinity for the PknB PASTAs. We do find a moderate effect of a high affinity peptide on resuscitation of dormant cells. While the PASTA domains of PknB may play a role in stimulating growth by binding exogenous peptidoglycan fragments, our data indicate that a major function of these domains is for proper PknB localization, likely through binding of peptidoglycan fragments produced locally at the mid-cell and the cell poles. These data suggest a model in which PknB is targeted to the sites of peptidoglycan turnover to regulate cell growth and cell division.

Vyšlo v časopise: The Extracytoplasmic Domain of the Ser/Thr Kinase PknB Binds Specific Muropeptides and Is Required for PknB Localization. PLoS Pathog 7(7): e32767. doi:10.1371/journal.ppat.1002182
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002182

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1. GoehringNWBeckwithJ 2005 Diverse paths to midcell: assembly of the bacterial cell division machinery. Curr Biol 15 R514 526

2. ErringtonJDanielRAScheffersDJ 2003 Cytokinesis in bacteria. Microbiol Mol Biol Rev 67 52 65

3. BlackmanSASmithTJFosterSJ 1998 The role of autolysins during vegetative growth of Bacillus subtilis 168. Microbiology 144 Pt 1 73 82

4. VollmerWBlanotDde PedroMA 2008 Peptidoglycan structure and architecture. FEMS Microbiol Rev 32 149 167

5. VollmerWJorisBCharlierPFosterS 2008 Bacterial peptidoglycan (murein) hydrolases. FEMS Microbiol Rev 32 259 286

6. MorlotCUeharaTMarquisKABernhardtTGRudnerDZ 2010 A highly coordinated cell wall degradation machine governs spore morphogenesis in Bacillus subtilis. Genes Dev 24 411 422

7. ChaoMCRubinEJ 2010 Letting sleeping dos lie: does dormancy play a role in tuberculosis? Annu Rev Microbiol 64 293 311

8. MukamolovaGVKaprelyantsASYoungDIYoungMKellDB 1998 A bacterial cytokine. Proc Natl Acad Sci U S A 95 8916 8921

9. CamusJCPryorMJMedigueCColeST 2002 Re-annotation of the genome sequence of Mycobacterium tuberculosis H37Rv. Microbiology 148 2967 2973

10. DowningKJMischenkoVVShleevaMOYoungDIYoungM 2005 Mutants of Mycobacterium tuberculosis lacking three of the five rpf-like genes are defective for growth in vivo and for resuscitation in vitro. Infect Immun 73 3038 3043

11. KanaBDGordhanBGDowningKJSungNVostroktunovaG 2008 The resuscitation-promoting factors of Mycobacterium tuberculosis are required for virulence and resuscitation from dormancy but are collectively dispensable for growth in vitro. Mol Microbiol 67 672 684

12. Russell-GoldmanEXuJWangXChanJTufarielloJM 2008 A Mycobacterium tuberculosis Rpf double-knockout strain exhibits profound defects in reactivation from chronic tuberculosis and innate immunity phenotypes. Infect Immun 76 4269 4281

13. MukamolovaGVMurzinAGSalinaEGDeminaGRKellDB 2006 Muralytic activity of Micrococcus luteus Rpf and its relationship to physiological activity in promoting bacterial growth and resuscitation. Mol Microbiol 59 84 98

14. YeatsCFinnRDBatemanA 2002 The PASTA domain: a beta-lactam-binding domain. Trends Biochem Sci 27 438 440

15. ShahIMLaaberkiMHPophamDLDworkinJ 2008 A eukaryotic-like Ser/Thr kinase signals bacteria to exit dormancy in response to peptidoglycan fragments. Cell 135 486 496

16. ColeSTBroschRParkhillJGarnierTChurcherC 1998 Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature 393 537 544

17. MukamolovaGVTurapovOAYoungDIKaprelyantsASKellDB 2002 A family of autocrine growth factors in Mycobacterium tuberculosis. Mol Microbiol 46 623 635

18. MukamolovaGVYanopolskayaNDKellDBKaprelyantsAS 1998 On resuscitation from the dormant state of Micrococcus luteus. Antonie Van Leeuwenhoek 73 237 243

19. MahapatraSCrickDCMcNeilMRBrennanPJ 2008 Unique structural features of the peptidoglycan of Mycobacterium leprae. J Bacteriol 190 655 661

20. LavollayMArthurMFourgeaudMDubostLMarieA 2008 The peptidoglycan of stationary-phase Mycobacterium tuberculosis predominantly contains cross-links generated by L,D-transpeptidation. J Bacteriol 190 4360 4366

21. LeeMHesekDShahIMOliverAGDworkinJ 2010 Synthetic peptidoglycan motifs for germination of bacterial spores. Chembiochem: a Chembiochem 11 2525 2529

22. KangCMNyayapathySLeeJYSuhJWHussonRN 2008 Wag31, a homologue of the cell division protein DivIVA, regulates growth, morphology and polar cell wall synthesis in mycobacteria. Microbiology 154 725 735

23. KangCMAbbottDWParkSTDascherCCCantleyLC 2005 The Mycobacterium tuberculosis serine/threonine kinases PknA and PknB: substrate identification and regulation of cell shape. Genes Dev 19 1692 1704

24. LamHOhDCCavaFTakacsCNClardyJ 2009 D-amino acids govern stationary phase cell wall remodeling in bacteria. Science 325 1552 1555

25. Kolodkin-GalIRomeroDCaoSClardyJKolterR 2010 D-amino acids trigger biofilm disassembly. Science 328 627 629

26. DessenAMouzNGordonEHopkinsJDidebergO 2001 Crystal structure of PBP2x from a highly penicillin-resistant Streptococcus pneumoniae clinical isolate: a mosaic framework containing 83 mutations. J Biol Chem 276 45106 45112

27. BarthePMukamolovaGVRoumestandCCohen-GonsaudM 2010 The structure of PknB extracellular PASTA domain from Mycobacterium tuberculosis suggests a ligand-dependent kinase activation. Structure 18 606 615

28. YoungTADelagoutteBEndrizziJAFalickAMAlberT 2003 Structure of Mycobacterium tuberculosis PknB supports a universal activation mechanism for Ser/Thr protein kinases. Nat Struct Biol 10 168 174

29. MieczkowskiCIavaroneATAlberT 2008 Auto-activation mechanism of the Mycobacterium tuberculosis PknB receptor Ser/Thr kinase. Embo J 27 3186 3197

30. Ortiz-LombardiaMPompeoFBoitelBAlzariPM 2003 Crystal structure of the catalytic domain of the PknB serine/threonine kinase from Mycobacterium tuberculosis. J Biol Chem 278 13094 13100

31. PrisicSDankwaSSchwartzDChouMFLocasaleJW 2010 Extensive phosphorylation with overlapping specificity by Mycobacterium tuberculosis serine/threonine protein kinases. Proc Natl Acad Sci U S A 107 7521 7526

32. DasguptaADattaPKunduMBasuJ 2006 The serine/threonine kinase PknB of Mycobacterium tuberculosis phosphorylates PBPA, a penicillin-binding protein required for cell division. Microbiology 152 493 504

33. SurekaKHossainTMukherjeePChatterjeePDattaP 2010 Novel role of phosphorylation-dependent interaction between FtsZ and FipA in mycobacterial cell division. PLoS ONE 5 e8590

34. ChowdhuryABoonsGJ 2005 The synthesis of diaminopimelic acid containing peptidoglycan fragments using metathesis cross coupling. Tetrahedron Lett 46 1675 1678

35. ChowdhuryASiriwardenaABoonsGJ 2002 A highly convergent approach for the synthesis of disaccharide repeating units of peptidoglycan. Tetrahedron Lett 43 7805 7807

36. GuanRRoychowdhuryAEmberBKumarSBoonsGJ 2004 Structural basis for peptidoglycan binding by peptidoglycan recognition proteins. Proc Natl Acad Sci U S A 101 17168 17173

37. KumarSRoychowdhuryAEmberBWangQGuanR 2005 Selective recognition of synthetic lysine and meso-diaminopimelic acid-type peptidoglycan fragments by human peptidoglycan recognition proteins Iα and S. J Biol Chem 280 37005 37012

38. RoychowdhuryAWolfertMABoonsGJ 2005 Synthesis and proinflammatory properties of muramyl tripeptides containing lysine and diaminopimelic acid moieties. Chembiochem 6 2088 2097

39. SwaminathanCPBrownPHRoychowdhuryAWangQGuanR 2006 Dual strategies for peptidoglycan discrimination by peptidoglycan recognition proteins (PGRPs). Proc Natl Acad Sci U S A 103 684 689

40. AsongJWolfertMAMaitiKKMillerDBoonsGJ 2009 Binding and cellular activation studies reveal that Toll-like receptor 2 can differentially recognize peptidoglycan from Gram-positive and Gram-negative bacteria. J Biol Chem 284 8643 8653

41. SunZZhangY 1999 Spent culture supernatant of Mycobacterium tuberculosis H37Ra improves viability of aged cultures of this strain and allows small inocula to initiate growth. J Bacteriol 181 7626 7628

42. de ManJ 1975 The probability of most probable numbers. Eur J Appl Microbiol 1 67 78