Self-Protection against Gliotoxin—A Component of the Gliotoxin Biosynthetic Cluster, GliT, Completely Protects Against Exogenous Gliotoxin
Gliotoxin, and other related molecules, are encoded by multi-gene clusters and biosynthesized by fungi using non-ribosomal biosynthetic mechanisms. Almost universally described in terms of its toxicity towards mammalian cells, gliotoxin has come to be considered as a component of the virulence arsenal of Aspergillus fumigatus. Here we show that deletion of a single gene, gliT, in the gliotoxin biosynthetic cluster of two A. fumigatus strains, rendered the organism highly sensitive to exogenous gliotoxin and completely disrupted gliotoxin secretion. Addition of glutathione to both A. fumigatus ΔgliT strains relieved gliotoxin inhibition. Moreover, expression of gliT appears to be independently regulated compared to all other cluster components and is up-regulated by exogenous gliotoxin presence, at both the transcript and protein level. Upon gliotoxin exposure, gliT is also expressed in A. fumigatus ΔgliZ, which cannot express any other genes in the gliotoxin biosynthetic cluster, indicating that gliT is primarily responsible for protecting this strain against exogenous gliotoxin. GliT exhibits a gliotoxin reductase activity up to 9 µM gliotoxin and appears to prevent irreversible depletion of intracellular glutathione stores by reduction of the oxidized form of gliotoxin. Cross-species resistance to exogenous gliotoxin is acquired by A. nidulans and Saccharomyces cerevisiae, respectively, when transformed with gliT. We hypothesise that the primary role of gliotoxin may be as an antioxidant and that in addition to GliT functionality, gliotoxin secretion may be a component of an auto-protective mechanism, deployed by A. fumigatus to protect itself against this potent biomolecule.
Vyšlo v časopise:
Self-Protection against Gliotoxin—A Component of the Gliotoxin Biosynthetic Cluster, GliT, Completely Protects Against Exogenous Gliotoxin. PLoS Pathog 6(6): e32767. doi:10.1371/journal.ppat.1000952
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1000952
Souhrn
Gliotoxin, and other related molecules, are encoded by multi-gene clusters and biosynthesized by fungi using non-ribosomal biosynthetic mechanisms. Almost universally described in terms of its toxicity towards mammalian cells, gliotoxin has come to be considered as a component of the virulence arsenal of Aspergillus fumigatus. Here we show that deletion of a single gene, gliT, in the gliotoxin biosynthetic cluster of two A. fumigatus strains, rendered the organism highly sensitive to exogenous gliotoxin and completely disrupted gliotoxin secretion. Addition of glutathione to both A. fumigatus ΔgliT strains relieved gliotoxin inhibition. Moreover, expression of gliT appears to be independently regulated compared to all other cluster components and is up-regulated by exogenous gliotoxin presence, at both the transcript and protein level. Upon gliotoxin exposure, gliT is also expressed in A. fumigatus ΔgliZ, which cannot express any other genes in the gliotoxin biosynthetic cluster, indicating that gliT is primarily responsible for protecting this strain against exogenous gliotoxin. GliT exhibits a gliotoxin reductase activity up to 9 µM gliotoxin and appears to prevent irreversible depletion of intracellular glutathione stores by reduction of the oxidized form of gliotoxin. Cross-species resistance to exogenous gliotoxin is acquired by A. nidulans and Saccharomyces cerevisiae, respectively, when transformed with gliT. We hypothesise that the primary role of gliotoxin may be as an antioxidant and that in addition to GliT functionality, gliotoxin secretion may be a component of an auto-protective mechanism, deployed by A. fumigatus to protect itself against this potent biomolecule.
Zdroje
1. GardinerDM
WaringP
HowlettBJ
2005 The epipolythiodioxopiperazine (ETP) class of fungal toxins: distribution, mode of action, functions and biosynthesis. Microbiology 151 1021 1032
2. Kwon-ChungKJ
SuguiJA
2008 What do we know about the role of gliotoxin in the pathobiology of Aspergillus fumigatus? Med Mycol 2 1 7
3. FoxEM
HowlettBJ
2008 Biosynthetic gene clusters for epipolythiodioxopiperazines in filamentous fungi. Mycol Res 112 162 169
4. HurneAM
ChaiCL
WaringP
2000 Inactivation of rabbit muscle creatine kinase by reversible formation of an internal disulfide bond induced by the fungal toxin gliotoxin. J Biol Chem 275 25202 25206
5. TsunawakiS
YoshidaLS
NishidaS
KobayashiT
ShimoyamaT
2004 Fungal metabolite gliotoxin inhibits assembly of the human respiratory burst NADPH oxidase. Infect Immun 72 3373 3382
6. NishidaS
YoshidaLS
ShimoyamaT
NunoiH
KobayashiT
2005 Fungal metabolite gliotoxin targets flavocytochrome b558 in the activation of the human neutrophil NADPH oxidase. Infect Immun 73 235 244
7. GardinerDM
CozijnsenAJ
WilsonLM
PedrasMS
HowlettBJ
2004 The sirodesmin biosynthetic gene cluster of the plant pathogenic fungus Leptosphaeria maculans. Mol Microbiol 53 1307 1318
8. GardinerDM
HowlettBJ
2005 Bioinformatic and expression analysis of the putative gliotoxin biosynthetic gene cluster of Aspergillus fumigatus. FEMS Microbiol Lett 248 241 248
9. CramerRAJr
GamcsikMP
BrookingRM
NajvarLK
KirkpatrickWR
2006 Disruption of a nonribosomal peptide synthetase in Aspergillus fumigatus eliminates gliotoxin production. Eukaryot Cell 5 972 980
10. KupfahlC
HeinekampT
GeginatG
RuppertT
HärtlA
2006 Deletion of the gliP gene of Aspergillus fumigatus results in loss of gliotoxin production but has no effect on virulence of the fungus in a low-dose mouse infection model. Mol Microbiol 62 292 302
11. SuguiJA
PardoJ
ChangYC
ZaremberKA
NardoneG
2007 Gliotoxin is a virulence factor of Aspergillus fumigatus: gliP deletion attenuates virulence in mice immunosuppressed with hydrocortisone. Eukaryot Cell 6 1562 1569
12. SpikesS
XuR
NguyenCK
ChamilosG
KontoyiannisDP
2008 Gliotoxin production in Aspergillus fumigatus contributes to host-specific differences in virulence. J Infect Dis 197 479 486
13. GardinerDM
JarvisRS
HowlettBJ
2005 The ABC transporter gene in the sirodesmin biosynthetic gene cluster of Leptosphaeria maculans is not essential for sirodesmin production but facilitates self-protection. Fungal Genet Biol 42 257 263
14. BokJW
ChungD
BalajeeSA
MarrKA
AndesD
2006 GliZ, a transcriptional regulator of gliotoxin biosynthesis, contributes to Aspergillus fumigatus virulence. Infect Immun 74 6761 6768
15. BalibarCJ
WalshCT
2006 GliP, a multimodular nonribosomal peptide synthetase in Aspergillus fumigatus, makes the diketopiperazine scaffold of gliotoxin. Biochemistry 45 15029 15038
16. SrinivasanU
BalaA
JaoSC
StarkeDW
JordanTW
2006 Selective inactivation of glutaredoxin by sporidesmin and other epidithiopiperazinediones. Biochemistry 45 8978 8987
17. BernardoPH
BraschN
ChaiCL
WaringP
2003 A novel redox mechanism for the glutathione-dependent reversible uptake of a fungal toxin in cells. J Biol Chem 278 46549 46555
18. ChoiHS
ShimJS
KimJA
KangSW
KwonHJ
2007 Discovery of gliotoxin as a new small molecule targeting thioredoxin redox system. Biochem Biophys Res Commun 359 523 528
19. ChamilosG
LewisRE
LamarisGA
AlbertND
KontoyiannisDP
2008 Genomewide screening for genes associated with gliotoxin resistance and sensitivity in Saccharomyces cerevisiae. Antimicrob Agents Chemother 52 1325 1329
20. NielsenML
AlbertsenL
LettierG
NielsenJB
MortensenUH
2006 Efficient PCR-based gene targeting with a recyclable marker for Aspergillus nidulans Fungal Genet Biol 43 54 64
21. KuboderaT
YamashitaN
NishimuraA
2000 Pyrithiamine resistance gene (ptrA) of Aspergillus oryzae: cloning, characterization and application as a dominant selectable marker for transformation. Biosci Biotechnol Biochem 64 1416 1421
22. PatronNJ
WallerRF
CozijnsenAJ
StraneyDC
GardinerDM
2007 Origin and distribution of epipolythiodioxopiperazine (ETP) gene clusters in filamentous ascomycetes. BMC Evol Biol 7 174
23. GalaganJE
CalvoSE
CuomoC
MaLJ
WortmanJR
2005 Sequencing of Aspergillus nidulans and comparative analysis with A. fumigatus and A. oryzae. Nature 438 1105 1115
24. Hjorth-SørensenB
HoffmannER
LissinNM
SewellAK
JakobsenBK
2001 Activation of heat shock transcription factor in yeast is not influenced by the levels of expression of heat shock proteins. Mol Microbiol 39 914 923
25. ThönM
Al-AbdallahQ
HortschanskyP
BrakhageAA
2007 The thioredoxin system of the filamentous fungus Aspergillus nidulans: impact on development and oxidative stress response. J Biol Chem 282 27259 27269
26. LeClerqueA
WanH
2007 Novel dominant selection marker for the transformation of fungi. US Patent 2007178594
27. ArcherDB
DyerPS
2004 From genomics to post-genomics in Aspergillus. Curr Opin Microbiol 7 499 504
28. Rodríguez-SáizM
LemboM
BertettiL
MuracaR
VelascoJ
2004 Strain improvement for cephalosporin production by Acremonium chrysogenum using geneticin as a suitable transformation marker. FEMS Microbiol Lett 235 43 49
29. LiX
KimSK
NamKW
KangJS
ChoiHD
2006 A new antibacterial dioxopiperazine alkaloid related to gliotoxin from a marine isolate of the fungus Pseudallescheria. J Antibiot (Tokyo) 59 248 250
30. LosadaL
AjayiO
FrisvadJC
YuJ
NiermanWC
2009 Effect of competition on the production and activity of secondary metabolites in Aspergillus species. Med Mycol 47 Suppl 1 S88 96
31. WatanabeA
KameiK
SekineT
WakuM
NishimuraK
2004 Effect of aeration on gliotoxin production by Aspergillus fumigatus in its culture filtrate. Mycopathologia 157 19 27
32. PontecorvoG
RroperJA
HemmonsLM
MacDonaldKD
BuftonAW
1953 The genetics of Aspergillus nidulans. Adv Genet 5 141 238
33. PuntPJ
OliverRP
DingemanseMA
PouwelsPH
van den HondelCA
1987 Transformation of Aspergillus based on the hygromycin B resistance marker from Escherichia coli. Gene 56 117 124
34. LangfelderK
PhilippeB
JahnB
LatgéJP
BrakhageAA
2001 Differential expression of the Aspergillus fumigatus pksP gene detected in vitro and in vivo with green fluorescent protein. Infect Immun 69 6411 6418
35. TilburnJ
Sánchez-FerreroJC
ReoyoE
ArstHNJr
PeñalvaMA
2005 Mutational analysis of the pH signal transduction component PalC of Aspergillus nidulans supports distant similarity to BRO1 domain family members. Genetics 171 393 401
36. CarberryS
NevilleCM
KavanaghKA
DoyleS
2006 Analysis of major intracellular proteins of Aspergillus fumigatus by MALDI mass spectrometry: identification and characterisation of an elongation factor 1B protein with glutathione transferase activity. Biochem Biophys Res Commun 341 1096 1104
37. ReevesEP
MessinaCG
DoyleS
KavanaghK
2004 Correlation between gliotoxin production and virulence of Aspergillus fumigatus in Galleria mellonella. Mycopathologia 158 73 79
38. BradfordMM
1976 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72 248 254
39. HillKE
McCollumGW
BurkRF
1997 Determination of thioredoxin reductase activity in rat liver supernatant. Anal Biochem 253 123 125
40. SpelligT
BottinA
KahmannR
1996 Green fluorescent protein (GFP) as a new vital marker in the phytopathogenic fungus Ustilago maydis. Mol Gen Genet 252 503 509
41. LooversHM
GuinanE
JonesGW
2007 Importance of Hsp70 ATPase domain in prion propagation. Genetics 175 621 630
42. SchwimmerC
Masison
DC
2002 Antagonistic interactions between yeast [PSI+] and [URE3] prions and curing of [URE3] by Hsp70 protein chaperone Ssa1p but not by Ssa2p. Mol Cell Biol 22 3590 3598
43. HearnVM
MackenzieDW
1980 Mycelial antigens from two strains of Aspergillus fumigatus: an analysis by two-dimensional immunoelectrophoresis. Mykosen 23 549 562
44. TaylorJJ
BurroughsEJ
1973 Experimental avian aspergillosis. Mycopathol Mycol Appl 51 131 141
45. ObereggerH
EisendleM
SchrettlM
GraessleS
HaasH
2003 4′-phosphopantetheinyl transferase-encoding npgA is essential for siderophore biosynthesis in Aspergillus nidulans. Curr Genet 44 211 215
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Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
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