#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Two Coregulated Efflux Transporters Modulate Intracellular Heme and Protoporphyrin IX Availability in


Streptococcus agalactiae is a major neonatal pathogen whose infectious route involves septicemia. This pathogen does not synthesize heme, but scavenges it from blood to activate a respiration metabolism, which increases bacterial cell density and is required for full virulence. Factors that regulate heme pools in S. agalactiae are unknown. Here we report that one main strategy of heme and protoporphyrin IX (PPIX) homeostasis in S. agalactiae is based on a regulated system of efflux using two newly characterized operons, gbs1753 gbs1752 (called pefA pefB), and gbs1402 gbs1401 gbs1400 (called pefR pefC pefD), where pef stands for ‘porphyrin-regulated efflux’. In vitro and in vivo data show that PefR, a MarR-superfamily protein, is a repressor of both operons. Heme or PPIX both alleviate PefR-mediated repression. We show that bacteria inactivated for both Pef efflux systems display accrued sensitivity to these porphyrins, and give evidence that they accumulate intracellularly. The ΔpefR mutant, in which both pef operons are up-regulated, is defective for heme-dependent respiration, and attenuated for virulence. We conclude that this new efflux regulon controls intracellular heme and PPIX availability in S. agalactiae, and is needed for its capacity to undergo respiration metabolism, and to infect the host.


Vyšlo v časopise: Two Coregulated Efflux Transporters Modulate Intracellular Heme and Protoporphyrin IX Availability in. PLoS Pathog 6(4): e32767. doi:10.1371/journal.ppat.1000860
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1000860

Souhrn

Streptococcus agalactiae is a major neonatal pathogen whose infectious route involves septicemia. This pathogen does not synthesize heme, but scavenges it from blood to activate a respiration metabolism, which increases bacterial cell density and is required for full virulence. Factors that regulate heme pools in S. agalactiae are unknown. Here we report that one main strategy of heme and protoporphyrin IX (PPIX) homeostasis in S. agalactiae is based on a regulated system of efflux using two newly characterized operons, gbs1753 gbs1752 (called pefA pefB), and gbs1402 gbs1401 gbs1400 (called pefR pefC pefD), where pef stands for ‘porphyrin-regulated efflux’. In vitro and in vivo data show that PefR, a MarR-superfamily protein, is a repressor of both operons. Heme or PPIX both alleviate PefR-mediated repression. We show that bacteria inactivated for both Pef efflux systems display accrued sensitivity to these porphyrins, and give evidence that they accumulate intracellularly. The ΔpefR mutant, in which both pef operons are up-regulated, is defective for heme-dependent respiration, and attenuated for virulence. We conclude that this new efflux regulon controls intracellular heme and PPIX availability in S. agalactiae, and is needed for its capacity to undergo respiration metabolism, and to infect the host.


Zdroje

1. MenseSM

ZhangL

2006 Heme: a versatile signaling molecule controlling the activities of diverse regulators ranging from transcription factors to MAP kinases. Cell Res 16 681 692

2. ShepherdM

HeathMD

PooleRK

2007 NikA binds heme: a new role for an Escherichia coli periplasmic nickel-binding protein. Biochemistry 46 5030 5037

3. Gilles-GonzalezMA

GonzalezG

2004 Signal transduction by heme-containing PAS-domain proteins. J Appl Physiol 96 774 783

4. KumarS

BandyopadhyayU

2005 Free heme toxicity and its detoxification systems in human. Toxicol Lett 157 175 188

5. LetoffeS

HeuckG

DelepelaireP

LangeN

WandersmanC

2009 Bacteria capture iron from heme by keeping tetrapyrrol skeleton intact. Proc Natl Acad Sci U S A

6. BaughnAD

MalamyMH

2004 The strict anaerobe Bacteroides fragilis grows in and benefits from nanomolar concentrations of oxygen. Nature 427 441 444

7. BrooijmansRJ

de VosWM

HugenholtzJ

2009 Lactobacillus plantarum WCFS1 electron transport chains. Appl Environ Microbiol 75 3580 3585

8. DuwatP

SouriceS

CesselinB

LamberetG

VidoK

2001 Respiration capacity of the fermenting bacterium Lactococcus lactis and its positive effects on growth and survival. J Bacteriol 183 4509 4516

9. GauduP

VidoK

CesselinB

KulakauskasS

TremblayJ

2002 Respiration capacity and consequences in Lactococcus lactis. Antonie Van Leeuwenhoek 82 263 269

10. HanYH

SmibertRM

KriegNR

1992 Cytochrome composition and oxygen-dependent respiration-driven proton translocation in Wolinella curva, Wolinella recta, Bacteroides ureolyticus, and Bacteroides gracilis. Can J Microbiol 38 104 110

11. HuyckeMM

MooreD

JoyceW

WiseP

ShepardL

2001 Extracellular superoxide production by Enterococcus faecalis requires demethylmenaquinone and is attenuated by functional terminal quinol oxidases. Mol Microbiol 42 729 740

12. PedersenMB

GarriguesC

TuphileK

BrunC

VidoK

2008 Impact of aeration and heme-activated respiration on Lactococcus lactis gene expression: identification of a heme-responsive operon. J Bacteriol 190 4903 4911

13. RezaikiL

CesselinB

YamamotoY

VidoK

van WestE

2004 Respiration metabolism reduces oxidative and acid stress to improve long-term survival of Lactococcus lactis. Mol Microbiol 53 1331 1342

14. RitcheyTW

SeeleyHW

1974 Cytochromes in Streptococcus faecalis var. zymogenes grown in a haematin-containing medium. J Gen Microbiol 85 220 228

15. SijpesteijnAK

1970 Induction of cytochrome formation and stimulation of oxidative dissimilation by hemin in Streptococcus lactis and Leuconostoc mesenteroides. Antonie Van Leeuwenhoek 36 335 348

16. WinstedtL

FrankenbergL

HederstedtL

von WachenfeldtC

2000 Enterococcus faecalis V583 contains a cytochrome bd-type respiratory oxidase. J Bacteriol 182 3863 3866

17. YamamotoY

PoyartC

Trieu-CuotP

LamberetG

GrussA

2005 Respiration metabolism of Group B Streptococcus is activated by environmental haem and quinone and contributes to virulence. Mol Microbiol 56 525 534

18. YamamotoY

PoyartC

Trieu-CuotP

LamberetG

GrussA

2006 Roles of environmental heme, and menaquinone, in Streptococcus agalactiae. Biometals 19 205 210

19. FrankenbergL

BrugnaM

HederstedtL

2002 Enterococcus faecalis heme-dependent catalase. J Bacteriol 184 6351 6356

20. AllenCE

SchmittMP

2009 HtaA is an iron-regulated hemin binding protein involved in the utilization of heme iron in Corynebacterium diphtheriae. J Bacteriol 191 2638 2648

21. AndrewsSC

RobinsonAK

Rodriguez-QuinonesF

2003 Bacterial iron homeostasis. FEMS Microbiol Rev 27 215 237

22. BibbLA

KunkleCA

SchmittMP

2007 The ChrA-ChrS and HrrA-HrrS signal transduction systems are required for activation of the hmuO promoter and repression of the hemA promoter in Corynebacterium diphtheriae. Infect Immun 75 2421 2431

23. CescauS

CwermanH

LetoffeS

DelepelaireP

WandersmanC

2007 Heme acquisition by hemophores. Biometals 20 603 613

24. KaurAP

LanskyIB

WilksA

2009 The role of the cytoplasmic heme-binding protein (PhuS) of Pseudomonas aeruginosa in intracellular heme trafficking and iron homeostasis. J Biol Chem 284 56 66

25. QiZ

HamzaI

O'BrianMR

1999 Heme is an effector molecule for iron-dependent degradation of the bacterial iron response regulator (Irr) protein. Proc Natl Acad Sci U S A 96 13056 13061

26. StauffDL

BagaleyD

TorresVJ

JoyceR

AndersonKL

2008 Staphylococcus aureus HrtA is an ATPase required for protection against heme toxicity and prevention of a transcriptional heme stress response. J Bacteriol 190 3588 3596

27. TorresVJ

StauffDL

PishchanyG

BezbradicaJS

GordyLE

2007 A Staphylococcus aureus regulatory system that responds to host heme and modulates virulence. Cell Host Microbe 1 109 119

28. StauffDL

SkaarEP

2009 Bacillus anthracis HssRS signaling to HrtAB regulates heme resistance during infection. Mol Microbiol

29. DoranKS

NizetV

2004 Molecular pathogenesis of neonatal group B streptococcal infection: no longer in its infancy. Mol Microbiol 54 23 31

30. LindahlG

Stalhammar-CarlemalmM

AreschougT

2005 Surface proteins of Streptococcus agalactiae and related proteins in other bacterial pathogens. Clin Microbiol Rev 18 102 127

31. PaoSS

PaulsenIT

SaierMHJr

1998 Major facilitator superfamily. Microbiol Mol Biol Rev 62 1 34

32. PoyartC

Trieu-CuotP

1997 A broad-host-range mobilizable shuttle vector for the construction of transcriptional fusions to beta-galactosidase in gram-positive bacteria. FEMS Microbiol Lett 156 193 198

33. WilkinsonSP

GroveA

2006 Ligand-responsive transcriptional regulation by members of the MarR family of winged helix proteins. Curr Issues Mol Biol 8 51 62

34. ProvidentiMA

WyndhamRC

2001 Identification and functional characterization of CbaR, a MarR-like modulator of the cbaABC-encoded chlorobenzoate catabolism pathway. Appl Environ Microbiol 67 3530 3541

35. WilkinsonSP

GroveA

2004 HucR, a novel uric acid-responsive member of the MarR family of transcriptional regulators from Deinococcus radiodurans. J Biol Chem 279 51442 51450

36. KaatzGW

DeMarcoCE

SeoSM

2006 MepR, a repressor of the Staphylococcus aureus MATE family multidrug efflux pump MepA, is a substrate-responsive regulatory protein. Antimicrob Agents Chemother 50 1276 1281

37. FriedmanDB

StauffDL

PishchanyG

WhitwellCW

TorresVJ

2006 Staphylococcus aureus redirects central metabolism to increase iron availability. PLoS Pathog 2 e87 doi:10.1371/journal.ppat.0020087

38. EpeB

PflaumM

HaringM

HeglerJ

RudigerH

1993 Use of repair endonucleases to characterize DNA damage induced by reactive oxygen species in cellular and cell-free systems. Toxicol Lett 67 57 72

39. MalikZ

LadanH

NitzanY

EhrenbergB

1990 The bactericidal activity of a deuteroporphyrin-hemin mixture on gram-positive bacteria. A microbiological and spectroscopic study. J Photochem Photobiol B 6 419 430

40. EllisonDW

MillerVL

2006 Regulation of virulence by members of the MarR/SlyA family. Curr Opin Microbiol 9 153 159

41. AlekshunMN

LevySB

MealyTR

SeatonBA

HeadJF

2001 The crystal structure of MarR, a regulator of multiple antibiotic resistance, at 2.3 A resolution. Nat Struct Biol 8 710 714

42. LimD

PooleK

StrynadkaNC

2002 Crystal structure of the MexR repressor of the mexRAB-oprM multidrug efflux operon of Pseudomonas aeruginosa. J Biol Chem 277 29253 29259

43. BrochetM

CouveE

ZouineM

VallaeysT

RusniokC

2006 Genomic diversity and evolution within the species Streptococcus agalactiae. Microbes Infect 8 1227 1243

44. LeighJA

1999 Streptococcus uberis: a permanent barrier to the control of bovine mastitis? Vet J 157 225 238

45. WardPN

HoldenMT

LeighJA

LennardN

BignellA

2009 Evidence for niche adaptation in the genome of the bovine pathogen Streptococcus uberis. BMC Genomics 10 54

46. GorisJ

KonstantinidisKT

KlappenbachJA

CoenyeT

VandammeP

2007 DNA-DNA hybridization values and their relationship to whole-genome sequence similarities. Int J Syst Evol Microbiol 57 81 91

47. FiorentinoG

RoncaR

CannioR

RossiM

BartolucciS

2007 MarR-like transcriptional regulator involved in detoxification of aromatic compounds in Sulfolobus solfataricus. J Bacteriol 189 7351 7360

48. KaatzGW

McAleeseF

SeoSM

2005 Multidrug resistance in Staphylococcus aureus due to overexpression of a novel multidrug and toxin extrusion (MATE) transport protein. Antimicrob Agents Chemother 49 1857 1864

49. LeeEH

Rouquette-LoughlinC

FolsterJP

ShaferWM

2003 FarR regulates the farAB-encoded efflux pump of Neisseria gonorrhoeae via an MtrR regulatory mechanism. J Bacteriol 185 7145 7152

50. PooleK

TetroK

ZhaoQ

NeshatS

HeinrichsDE

1996 Expression of the multidrug resistance operon mexA-mexB-oprM in Pseudomonas aeruginosa: mexR encodes a regulator of operon expression. Antimicrob Agents Chemother 40 2021 2028

51. SeoaneAS

LevySB

1995 Characterization of MarR, the repressor of the multiple antibiotic resistance (mar) operon in Escherichia coli. J Bacteriol 177 3414 3419

52. KeelSB

DotyRT

YangZ

QuigleyJG

ChenJ

2008 A heme export protein is required for red blood cell differentiation and iron homeostasis. Science 319 825 828

53. QuigleyJG

YangZ

WorthingtonMT

PhillipsJD

SaboKM

2004 Identification of a human heme exporter that is essential for erythropoiesis. Cell 118 757 766

54. KrishnamurthyP

RossDD

NakanishiT

Bailey-DellK

ZhouS

2004 The stem cell marker Bcrp/ABCG2 enhances hypoxic cell survival through interactions with heme. J Biol Chem 279 24218 24225

55. KrishnamurthyP

XieT

SchuetzJD

2007 The role of transporters in cellular heme and porphyrin homeostasis. Pharmacol Ther 114 345 358

56. TatsumiR

WachiM

2008 TolC-dependent exclusion of porphyrins in Escherichia coli. J Bacteriol 190 6228 6233

57. BlankLM

KoebmannBJ

MichelsenO

NielsenLK

JensenPR

2001 Hemin reconstitutes proton extrusion in an H(+)-ATPase-negative mutant of Lactococcus lactis. J Bacteriol 183 6707 6709

58. BrooijmansRJ

PoolmanB

Schuurman-WoltersGK

de VosWM

HugenholtzJ

2007 Generation of a membrane potential by Lactococcus lactis through aerobic electron transport. J Bacteriol 189 5203 5209

59. KleinAH

ShullaA

ReimannSA

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

60. KoebmannB

BlankLM

SolemC

PetranovicD

NielsenLK

2008 Increased biomass yield of Lactococcus lactis during energetically limited growth and respiratory conditions. Biotechnol Appl Biochem 50 25 33

61. WaySS

SallustioS

MagliozzoRS

GoldbergMB

1999 Impact of either elevated or decreased levels of cytochrome bd expression on Shigella flexneri virulence. J Bacteriol 181 1229 1237

62. EndleyS

McMurrayD

FichtTA

2001 Interruption of the cydB locus in Brucella abortus attenuates intracellular survival and virulence in the mouse model of infection. J Bacteriol 183 2454 2462

63. ProctorRA

von EiffC

KahlBC

BeckerK

McNamaraP

2006 Small colony variants: a pathogenic form of bacteria that facilitates persistent and recurrent infections. Nat Rev Microbiol 4 295 305

64. ShiL

SohaskeyCD

KanaBD

DawesS

NorthRJ

2005 Changes in energy metabolism of Mycobacterium tuberculosis in mouse lung and under in vitro conditions affecting aerobic respiration. Proc Natl Acad Sci U S A 102 15629 15634

65. GaillotO

PoyartC

BercheP

Trieu-CuotP

1997 Molecular characterization and expression analysis of the superoxide dismutase gene from Streptococcus agalactiae. Gene 204 213 218

66. LamyMC

ZouineM

FertJ

VergassolaM

CouveE

2004 CovS/CovR of group B streptococcus: a two-component global regulatory system involved in virulence. Mol Microbiol 54 1250 1268

67. ChomczynskiP

SacchiN

1987 Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162 156 159

68. SambrookJ

FritschE

ManiatisT

1989 Molecular Cloning: A Laboratory Manual Cold Spring Harbor, New York Cold Spring Harbor Laboratory Press

69. MaguinE

PrevostH

EhrlichSD

GrussA

1996 Efficient insertional mutagenesis in lactococci and other gram-positive bacteria. J Bacteriol 178 931 935

70. BiswasI

GrussA

EhrlichSD

MaguinE

1993 High-efficiency gene inactivation and replacement system for gram-positive bacteria. J Bacteriol 175 3628 3635

71. LawJ

BuistG

HaandrikmanA

KokJ

VenemaG

1995 A system to generate chromosomal mutations in Lactococcus lactis which allows fast analysis of targeted genes. J Bacteriol 177 7011 7018

72. el KarouiM

EhrlichD

GrussA

1998 Identification of the lactococcal exonuclease/recombinase and its modulation by the putative Chi sequence. Proc Natl Acad Sci U S A 95 626 631

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2010 Číslo 4
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Aktuální možnosti diagnostiky a léčby litiáz
nový kurz
Autori: MUDr. Tomáš Ürge, PhD.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#