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 LaboratóriumČlánok vyšiel v časopise
PLOS Pathogens
2010 Číslo 4
- Očkování proti virové hemoragické horečce Ebola experimentální vakcínou rVSVDG-ZEBOV-GP
- Parazitičtí červi v terapii Crohnovy choroby a dalších zánětlivých autoimunitních onemocnění
- Koronavirus hýbe světem: Víte jak se chránit a jak postupovat v případě podezření?
Najčítanejšie v tomto čísle
- The Effect of Vaccination on the Evolution and Population Dynamics of Avian Paramyxovirus-1
- Reconstitution of SARS-Coronavirus mRNA Cap Methylation
- Deficiencies in Jasmonate-Mediated Plant Defense Reveal Quantitative Variation in Pathogenesis
- A Timescale for Evolution, Population Expansion, and Spatial Spread of an Emerging Clone of Methicillin-Resistant