Role of Acetyl-Phosphate in Activation of the Rrp2-RpoN-RpoS Pathway in

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Borrelia burgdorferi, the Lyme disease spirochete, dramatically alters its transcriptome and proteome as it cycles between the arthropod vector and mammalian host. During this enzootic cycle, a novel regulatory network, the Rrp2-RpoN-RpoS pathway (also known as the σ⁵⁴–σ^S sigma factor cascade), plays a central role in modulating the differential expression of more than 10% of all B. burgdorferi genes, including the major virulence genes ospA and ospC. However, the mechanism(s) by which the upstream activator and response regulator Rrp2 is activated remains unclear. Here, we show that none of the histidine kinases present in the B. burgdorferi genome are required for the activation of Rrp2. Instead, we present biochemical and genetic evidence that supports the hypothesis that activation of the Rrp2-RpoN-RpoS pathway occurs via the small, high-energy, phosphoryl-donor acetyl phosphate (acetyl∼P), the intermediate of the Ack-Pta (acetate kinase-phosphate acetyltransferase) pathway that converts acetate to acetyl-CoA. Supplementation of the growth medium with acetate induced activation of the Rrp2-RpoN-RpoS pathway in a dose-dependent manner. Conversely, the overexpression of Pta virtually abolished acetate-induced activation of this pathway, suggesting that acetate works through acetyl∼P. Overexpression of Pta also greatly inhibited temperature and cell density-induced activation of RpoS and OspC, suggesting that these environmental cues affect the Rrp2-RpoN-RpoS pathway by influencing acetyl∼P. Finally, overexpression of Pta partially reduced infectivity of B. burgdorferi in mice. Taken together, these findings suggest that acetyl∼P is one of the key activating molecule for the activation of the Rrp2-RpoN-RpoS pathway and support the emerging concept that acetyl∼P can serve as a global signal in bacterial pathogenesis.

Vyšlo v časopise: Role of Acetyl-Phosphate in Activation of the Rrp2-RpoN-RpoS Pathway in. PLoS Pathog 6(9): e32767. doi:10.1371/journal.ppat.1001104
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1001104

Souhrn

Zdroje

1. LaneRS

PiesmanJ

BurgdorferW

1991 Lyme borreliosis: relation of its causative agent to its vectors and hosts in North America and Europe. Annu Rev Entomol 36 587 609

2. CaimanoMJ

IyerR

EggersCH

GonzalezC

MortonEA

2007 Analysis of the RpoS regulon in Borrelia burgdorferi in response to mammalian host signals provides insight into RpoS function during the enzootic cycle. Mol Microbiol 65 1193 1217

3. FisherMA

GrimmD

HenionAK

EliasAF

StewartPE

2005 Borrelia burgdorferi σ54 is required for mammalian infection and vector transmission but not for tick colonization. Proc Natl Acad Sci U S A 102 5162 5167

4. OuyangZ

BlevinsJS

NorgardMV

2008 Transcriptional interplay among the regulators Rrp2, RpoN, and RpoS in Borrelia burgdorferi. Microbiology 154 2641 2658

5. CaimanoMJ

EggersCH

HazlettKRO

RadolfJD

2004 RpoS is not central to the general stress response in Borrelia burgdorferi but does control expression of one or more essential virulence determinants. Infect Immun 72 6433 6445

6. BurtnickMN

DowneyJS

BrettPJ

BoylanJA

FryeJG

2007 Insights into the complex regulation of rpoS in Borrelia burgdorferi. Mol Microbiol 65 277 293

7. LybeckerMC

SamuelsDS

2007 Temperature-induced regulation of RpoS by a small RNA in Borrelia burgdorferi. Mol Microbiol 64 1075 1089

8. HübnerA

YangX

NolenDM

PopovaTG

CabelloFC

2001 Expression of Borrelia burgdorferi OspC and DbpA is controlled by a RpoN-RpoS regulatory pathway. Proc Natl Acad Sci U S A 98 12724 12729

9. YangXF

PalU

AlaniSM

FikrigE

NorgardMV

2004 Essential role for OspA/B in the life cycle of the Lyme disease spirochete. J Exp Med 199 641 648

10. GrimmD

TillyK

ByramR

StewartPE

KrumJG

2004 Outer-surface protein C of the Lyme disease spirochete: A protein induced in ticks for infection of mammals. Proc Natl Acad Sci U S A 101 3142 3147

11. PalU

YangX

ChenM

BockenstedtLK

AndersonJF

2004 OspC facilitates Borrelia burgdorferi invasion of Ixodes scapularis salivary glands. J Clin Invest 113 220 230

12. TillyK

KrumJG

BestorA

JewettMW

GrimmD

2006 Borrelia burgdorferi OspC protein required exclusively in a crucial early stage of mammalian infection. Infect Immun 74 3554 3564

13. BattistiJM

BonoJL

RosaPA

SchrumpfME

SchwanTG

2008 Outer Surface Protein A Protects Lyme Disease Spirochetes from Acquired Host Immunity in the Tick Vector. Infect Immun 76 5228 5237

14. SmithAH

BlevinsJS

BachlaniGN

YangXF

NorgardMV

2007 Evidence that RpoS (σS) in Borrelia burgdorferi is controlled directly by RpoN (σ54/σN). J Bacteriol 189 2139 2144

15. BlevinsJS

XuH

HeM

NorgardMV

ReitzerL

2009 Rrp2, a σ54-dependent transcriptional activator of Borrelia burgdorferi, activates rpoS in an enhancer-independent manner. J Bacteriol 191 2902 2905

16. StudholmeDJ

BuckM

2000 The biology of enhancer-dependent transcriptional regulation in bacteria: insights from genome sequences. FEMS Microbiol Letts 186 1 9

17. FraserCM

CasjensS

HuangWM

SuttonGG

ClaytonR

1997 Genomic sequence of a Lyme disease spirochaete, Borrelia burgdorferi. Nature 390 580 586

18. YangXF

AlaniSM

NorgardMV

2003 The response regulator Rrp2 is essential for the expression of major membrane lipoproteins in Borrelia burgdorferi. Proc Natl Acad Sci U S A 100 11001 11006

19. NorthAK

KloseKE

StedmanKM

KustuS

1993 Prokaryotic enhancer-binding proteins reflect eukaryote-like modularity: the puzzle of nitrogen regulatory protein C. J Bacteriol 175 4267 4273

20. BoardmanBK

HeM

OuyangZ

XuH

PangX

2008 Essential role of the response regulator Rrp2 in the infectious cycle of Borrelia burgdorferi. Infect Immun 76 3844 3853

21. HochJA

2000 Two-component and phosphorelay signal transduction. Curr Opin Microbiol 3 165 170

22. MascherT

HelmannJD

UndenG

2006 Stimulus Perception in Bacterial Signal-Transducing Histidine Kinases. Microbiol Mol Biol Rev 70 910 938

23. StockAM

RobinsonVL

GoudreauPN

2000 Two component signal transduction. Annu Rev Biochem 69 183 215

24. YamamotoK

HiraoK

OshimaT

AibaH

UtsumiR

2005 Functional characterization in vitro of all twocomponent signal transduction systems from Escherichia coli. J Biol Chem 280 1448

25. NinfaAJ

NinfaEG

LupasAN

StockA

MagasanikB

1988 Crosstalk between bacterial chemotaxis signal transduction proteins and regulators of transcription of the Ntr regulon: evidence that nitrogen assimilation and chemotaxis are controlled by a common phosphotransfer mechanism. Proc Natl Acad Sci U S A 85 5492 5496

26. BijlsmaJE

GroismanEA

2003 Making informed decisions: regulatory interactions between two-component systems. Trends Micro 11 359 366

27. LaubMT

GoulianM

2007 Specificity in two-component signal transduction pathways. Annu Rev Genet 41 121 145

28. WannerBL

1992 Is cross regulation by phosphorylation of two-component response regulator proteins important in bacteria? J Bacteriol 174 2053 2058

29. McClearyWR

StockJB

NinfaAJ

1993 Is acetyl phosphate a global signal in Escherichia coli? J Bacteriol 175 2793 2798

30. LukatGS

McClearyWR

StockAM

StockJB

1992 Phosphorylation of bacterial response regulator proteins by low molecular weight phospho-donors. Proc Natl Acad Sci U S A 89 718 722

31. WannerBL

1993 Gene regulation by phosphate in enteric bacteria. J Cell Biochem 51 47 51

32. WolfeAJ

2005 The acetate switch. Microbiol Mol Biol Rev 69 12 50

33. WolfeAJ

2010 Physiologically relevant small phosphodonors link metabolism to signal transduction. Current Opinion in Microbiology 13 204 209

34. RogersEA

TerekhovaD

ZhangH

HovisKM

SchwartzI

2009 Rrp1, a cyclic-di-GMP-producing response regulator, is an important regulator of Borrelia burgdorferi core cellular functions. Mol Microbiol

35. EliasAF

StewartPE

GrimmD

CaimanoMJ

EggersCH

2002 Clonal Polymorphism of Borrelia burgdorferi Strain B31 MI: Implications for Mutagenesis in an Infectious Strain Background. Infect Immun 70 2139 2150

36. GilbertMA

MortonEA

BundleSF

SamuelsDS

2007 Artificial regulation of ospC expression in Borrelia burgdorferi Mol Microbiol 63 1259 1273

37. IndestKJ

RamamoorthyR

SoleM

GilmoreRD

JohnsonBJB

1997 Cell-density-dependent expression of Borrelia burgdorferi lipoproteins in vitro. Infect Immun 65 1165 1171

38. YangX

GoldbergMS

PopovaTG

SchoelerGB

WikelSK

2000 Interdependence of environmental factors influencing reciprocal patterns of gene expression in virulent Borrelia burgdorferi. Mol Microbiol 37 1470 1479

39. SchwanTG

PiesmanJ

GoldeWT

DolanMC

RosaPA

1995 Induction of an outer surface protein on Borrelia burgdorferi during tick feeding. Proc Natl Acad Sci U S A 92 2909 2913

40. AkinsDR

BourellKW

CaimanoMJ

NorgardMV

RadolfJD

1998 A new animal model for studying Lyme disease spirochetes in a mammalian host-adapted state. J Clin Invest 101 2240 2250

41. CaimanoMJ

EggersCH

GonzalezCA

RadolfJD

2005 Alternate sigma factor RpoS is required for the in vivo-specific repression of Borrelia burgdorferi plasmid lp54-borne ospA and lp6.6 genes. J Bacteriol 187 7845 7852

42. CaimanoM

2005 Cultivation of Borrelia burgdorferi in dialysis membrane chambers in rat peritonea. Curr Protoc Microbiol 12. Unit 12C.3

43. CharonNW

GoldsteinSF

2002 Genetics of motility and chemotaxis of a fascinating gruop of bacteria: the spirochetes. Annu Rev Genet 36 47 73

44. LiC

BakkerRG

MotalebMA

SartakovaML

CabelloFC

CharonNW

2002 Asymetrical flagellar rotation in Borrelia burgdorferi nonchemotactic mutants. Proc Natl Acad Sci USA 99 6169 6174

45. CarrollJA

GaronCF

SchwanTG

1999 Effects of environmental pH on membrane proteins in Borrelia burgdorferi. Infect Immun 67 3181 3187

46. ScharJ

SickmannA

BeierD

2005 Phosphorylation-Independent Activity of Atypical Response Regulators of Helicobacter pylori. J Bacteriol 187 3100 3109

47. RuizD

SalinasP

Lopez-RedondoML

CayuelaML

MarinaA

ContrerasA

2008 Phosphorylation-independent activation of the atypical response regulator NblR. Microbiology 154 3002 3015

48. FraserJS

MerlieJPJ

EcholsN

WeisfieldSR

MignotT

WemmerDE

ZusmanDR

AlberT

2007 An atypical receiver domain controls the dynamic polar localization of the Myxococcus xanthus social motility protein FrzS. Mol Microbiol 65 319 332

49. KleinAH

ShullaA

ReimannA

KeatingDH

WolfeAJ

2007 The intracellular concentration of acetyl phosphate in Escherichia coli is sufficient for direct phosphorylation of two-component response regulators. J Bacteriol 189 5574 5581

50. StevensonB

SchwanTG

RosaPA

1995 Temperature-related differential expression of antigens in the Lyme disease spirochete, Borrelia burgdorferi. Infect Immun 63 4535 4539

51. HydeJA

TrzeciakowskiJP

SkareJT

2007 Borrelia burgdorferi alters its gene expression and antigenic profile in response to CO2 levels. J Bacteriol 189 437 445

52. PrussBM

WolfeAJ

1994 Regulation of acetyl phosphate synthesis and degradation, and the control of flagellar expression in Escherichia coli. Mol Microbiol 12 973 984

53. HydeJA

ShawDK

SmithR

III

TrzeciakowskiJP

SkareJT

2010 Characterization of a conditional bosR mutant in Borrelia burgdorferi. Infect Immun 78 265 74

54. OuyangZ

KumarM

KariuT

HaqS

GoldbergM

2009 BosR (BB0647) governs virulence expression in Borrelia burgdorferi. Mol Microbiol 74 1331 1343

55. HydeJA

ShawDK

SmithLR

TrzeciakowskiJP

SkareJT

2009 The BosR regulatory protein of Borrelia burgdorferi interfaces with the RpoS regulatory pathway and modulates both the oxidative stress response and pathogenic properties of the Lyme disease spirochete. Mol Microbiol 74 1344 1355

56. SamuelsDS

RadolfJD

2009 Who is the BosR around here anyway? Mol Microbiol 74 1295 1299

57. de SilvaAM

FikrigE

1995 Growth and migration of Borrelia burgdorferi in Ixodes ticks during blood feeding. Am J Trop Med Hyg 53 397 404

58. PiesmanJ

SchneiderBS

ZeidnerNS

2001 Use of quantitative PCR to measure density of Borrelia burgdorferi in the midgut and salivary glands of feeding tick vectors. J Clin Microbiol 39 4145 4148

59. XuQ

McShanK

LiangFT

2007 Identification of an ospC operator critical for immune evasion of Borrelia burgdorferi. Mol Microbiol 64 220 231

60. SamuelsDS

1995 Electrotransformation of the spirochete Borrelia burgdorferi.

NickoloffJA

Methods in molecular biology. 45 ed 253-259 Totowa, NJ Humana Press

61. Labandeira-ReyM

SkareJT

2001 Decreased infectivity in Borrelia burgdorferi strain B31 is associated with loss of linear plasmid 25 or 28-1. Infect Immun 69 446 455

62. McDowellJV

SungSY

Labandeira-ReyM

SkareJT

MarconiRT

2001 Analysis of mechanisms associated with loss of infectivity of clonal populations of Borrelia burgdorferi B31MI. Infect Immun 69 3670 3677

63. PurserJE

NorrisSJ

2000 Correlation between plasmid content and infectivity in Borrelia burgdorferi. Proc Natl Acad Sci U S A 97 13865 13870

64. StewartPE

HoffJ

FischerE

KrumJG

RosaPA

2004 Genome-Wide Transposon Mutagenesis of Borrelia burgdorferi for Identification of Phenotypic Mutants. Appl Environ Microbiol 70 5973 5979

65. NorrisSJ

HowellJK

GarzaSA

FerdowsMS

BarbourAG

1995 High- and low-infectivity phenotypes of clonal populations of in vitro- cultured Borrelia burgdorferi. Infect Immun 63 2206 2212

66. YangX

PopovaTG

HagmanKE

WikelSK

SchoelerGB

1999 Identification, characterization, and expression of three new members of the Borrelia burgdorferi Mlp (2.9) lipoprotein gene family. Infect Immun 67 6008 6018

67. XuH

HeM

PangX

XuZC

PiesmanJ

YangXF

2009 Characterization of the highly regulated antigen BBA05 in the enzootic cycle of Borrelia burgdorferi. Infect Immun 15 1872 1887

68. QuonKC

MarczynskiGT

ShapiroL

1996 Cell Cycle Control by an Essential Bacterial Two-Component Signal Transduction Protein. Cell 84 83 93