Tri6 Is a Global Transcription Regulator in the Phytopathogen
In F.
graminearum, the transcriptional regulator Tri6 is encoded within the trichothecene gene cluster and regulates genes involved in the biosynthesis of the secondary metabolite deoxynivalenol (DON). The Tri6 protein with its Cys2His2 zinc-finger may also conform to the class of global transcription regulators. This class of global transcriptional regulators mediate various environmental cues and generally responds to the demands of cellular metabolism. To address this issue directly, we sought to find gene targets of Tri6 in F. graminearum grown in optimal nutrient conditions. Chromatin immunoprecipitation followed by Illumina sequencing (ChIP-Seq) revealed that in addition to identifying six genes within the trichothecene gene cluster, Tri1, Tri3, Tri6, Tri7, Tri12 and Tri14, the ChIP-Seq also identified 192 additional targets potentially regulated by Tri6. Functional classification revealed that, among the annotated genes, ∼40% are associated with cellular metabolism and transport and the rest of the target genes fall into the category of signal transduction and gene expression regulation. ChIP-Seq data also revealed Tri6 has the highest affinity toward its own promoter, suggesting that this gene could be subject to self-regulation. Electro mobility shift assays (EMSA) performed on the promoter of Tri6 with purified Tri6 protein identified a minimum binding motif of GTGA repeats as a consensus sequence. Finally, expression profiling of F. graminearum grown under nitrogen-limiting conditions revealed that 49 out of 198 target genes are differentially regulated by Tri6. The identification of potential new targets together with deciphering novel binding sites for Tri6, casts new light into the role of this transcriptional regulator in the overall growth and development of F. graminearum.
Vyšlo v časopise:
Tri6 Is a Global Transcription Regulator in the Phytopathogen. PLoS Pathog 7(9): e32767. doi:10.1371/journal.ppat.1002266
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1002266
Souhrn
In F.
graminearum, the transcriptional regulator Tri6 is encoded within the trichothecene gene cluster and regulates genes involved in the biosynthesis of the secondary metabolite deoxynivalenol (DON). The Tri6 protein with its Cys2His2 zinc-finger may also conform to the class of global transcription regulators. This class of global transcriptional regulators mediate various environmental cues and generally responds to the demands of cellular metabolism. To address this issue directly, we sought to find gene targets of Tri6 in F. graminearum grown in optimal nutrient conditions. Chromatin immunoprecipitation followed by Illumina sequencing (ChIP-Seq) revealed that in addition to identifying six genes within the trichothecene gene cluster, Tri1, Tri3, Tri6, Tri7, Tri12 and Tri14, the ChIP-Seq also identified 192 additional targets potentially regulated by Tri6. Functional classification revealed that, among the annotated genes, ∼40% are associated with cellular metabolism and transport and the rest of the target genes fall into the category of signal transduction and gene expression regulation. ChIP-Seq data also revealed Tri6 has the highest affinity toward its own promoter, suggesting that this gene could be subject to self-regulation. Electro mobility shift assays (EMSA) performed on the promoter of Tri6 with purified Tri6 protein identified a minimum binding motif of GTGA repeats as a consensus sequence. Finally, expression profiling of F. graminearum grown under nitrogen-limiting conditions revealed that 49 out of 198 target genes are differentially regulated by Tri6. The identification of potential new targets together with deciphering novel binding sites for Tri6, casts new light into the role of this transcriptional regulator in the overall growth and development of F. graminearum.
Zdroje
1. KangZBuchenauerH 2002 Studies on the infection process of Fusarium culmorum in wheat spikes: Degradation of host cell wall components and localization of trichothecene toxins in infected tissue. Eur J Plant Pathol 108 653 660
2. BennettJWKlichM 2003 Mycotoxins. Clinical Micro Reviews 16 497 516
3. PestkaJJSmolinskiAT 2005 Deoxynivalenol: toxicology and potential effects on humans. J Toxicol Environ Health B Critical Review 9 39 69
4. CalvoAMWilsonRABokJWKellerNP 2002 Relationship between secondary metabolism and fungal development. Microbiol and Mol Bio Reviews 66 447 459
5. GardinerDMKazanKMannersJM 2009 Nutrient profiling reveals potent inducers of trichothecene biosynthesis in fusarium graminearum. Fungal Genet Biol 46 604 613
6. YuJ-HKellerN 2005 Regulation of secondary metabolism in filamentous fungi. Annu Rev Phytopathol 43 437 58
7. GardinerDMOsborneSKazanKMannersJM 2009 Low pH regulates the production of deoxynivalenol by Fusarium graminearum. Microbiology 155 3149 3156
8. JiaoFKawakamiANakajimaT 2008 Effects of different carbon sources on trichothecene production and Tri gene expression by Fusarium graminearum in liquid culture. FEMS Microbiol Lett 285 212 219
9. OchiaiNTokaiTTakahashi-AndoNFujimuraMKimuraM 2007 Genetically engineered Fusarium as a tool to evaluate the effects of environmental factors on initiation of trichothecene biosynthesis. FEMS Microbiol Lett 275 53 61
10. HicksJYuJ-HKellerNAdamsTH 1997 Aspergillus sporulation and mycotoxin production both require inactivation of FadA G α protein-dependent signaling pathway. EMBO J 16 4916 4923
11. RozeLVBeudryRMKellerNPLinzJE 2004 Regulation of aflatoxin synthesis by FadA/cAMP/protein kinase A signaling in Aspergillus parasiticus. Mycopathologia 158 219 32
12. TagAHicksJGarifullinaGAkeCjrPhillipsTD 2000 G-protein signaling mediates differential production of toxic secondary metabolites. Mol Microbiol 38 658 665
13. PeñalvaMAArstHN 2004 Recent advances in the characterization of ambient pH regulation of gene expression in filamentous fungi and yeasts. Annu Rev Microbiol 58 425 451
14. MerhejJRichard-ForgetFBarreauC 2011 The pH regulatory factor Pac1 regulates Tri gene expression and trichothecene production in Fusarium graminearum. Fungal Genet Biol 48 275 284
15. FernandesMKellerNAdamsTH 1998 Sequence-specific binding by Aspergillus nidulans AflR, a C6 zinc cluster protein regulating mycotoxin biosynthesis. Mol Microbiol 28 1355 1365
16. Kye-YSeongPasqualiMHilburnKLMcCormickSPXuJ 2009 Global gene regulation by Fusarium transcription factors Tri6 and Tri10 reveals adaptations for toxin biosynthesis. Mol Microbiol 72 354 367
17. HohnTMKrishnaRProctorRH 1999 Characterization of a transcriptional activator controlling trichothecene toxin biosynthesis. Fungal Genet Biol 26 224 235
18. ProctorRHHohnTMMcCormickSPDesjardinsAE 1995 Tri6 encodes an unusual zinc finger protein involved in regulation of trichothecene biosynthesis in Fusarium sporotrichioides. Appl Environ Microbiol 61 1923 1930
19. PeplowAWTagAGGarifullinaGFBeremandMN 2003 Identification of new genes positively regulated by Tri10 and a regulatory network for trichothecene mycotoxin production. Appl Environ Microbiol 69 2731 2736
20. GardinerDMKazanKMannersJM 2009 Novel genes of Fusarium graminearum that negatively regulate deoxynivalenol production and virulence. Mol Plant-Microbe Interact 12 1588 1600
21. SchreiberKNasmithCAllardGSinghJSubramaniamR 2011 Found in translation: High-throughput chemical screening in Arabidopsis thaliana identifies small molecules that reduce Fusarium head blight disease in wheat. Mol Plant-Microbe Interact 24 640 648
22. JothiRCuddapahSBarskiACiuKZhaoK 2008 Genome-wide identification of in vivo protein-DNA binding sites from ChIP-Seq data. Nucleic Acids Res 36 5221 5231
23. SlaughterJC 1988 Nitrogen metabolism. BerryDR Physiology of Industrial Fungi Blackwell Scientific Publications 58 76
24. MarzlufGA 1997 Genetic regulation of nitrogen metabolism in the fungi. Microbiol Mol Biol Rev 61 17 32
25. BeopoulosAMrozovaZTheveniauFLe DallM-THapalaI 2008 Mastering lipid accumulation in the yeast Yarrowia lipolytica. Appl Env Microbiol 74 7779 7789
26. StammersDKRenJLeslieKNicholsCELambHK 2001 The structure of the negative transcriptional regulator NmrA reveals a structural superfamily which includes the short-chain dehydrogenase/reductases. The EMBO Journal 20 6619 6626
27. AndrianopoulosAKourambasSSharpJADavisMAHynesMJ 1998 Characterization of the Aspergillus nidulans nmrA gene involved in nitrogen metabolite repression. J Bacteriol 180 1973 1977
28. BluhmBHZhouXFlahertyJEXuJDunkleLD 2007 RAS2 regulates growth and pathogenesis in Fusarium graminearum. Mol Plant Microbe Interact 20 627 636
29. HoffmanCS 2005 Glucose sensing via the protein kinase A pathway in Schizosaccharomyces pombe. Biochem Soc Trans 33 257 260
30. FroeligerEHCarpenterBE 1996 NUT1, a major nitrogen regulatory gene in Magnaporthe grisea, is dispensable for pathogenicity. Mol Gen Genet 251 647 56
31. WongKHHynesMJDavisMA 2008 Recent advances in nitrogen regulation: A comparison between Saccharomyces cerevisiae and filamentous fungi. Eukaryotic Cell 7 917 925
32. MihlanMHomannVLiuT-WDTudzynskiB 2003 AREA directly mediates nitrogen regulation of gibberellin biosynthesis in Gibberella fujikuroi, but its activity is not affected by NMR. Mol Microbiol 47 975 991
33. WilsonRAArstrHNJr 1998 Mutational analysis of AREA, a transcriptional activator mediating nitrogen metabolite repression in Aspergillus nidulans and a member of the “streetwise” GATA family. Microbiol Mol Biol Reviews 62 586 596
34. SchönigBBrownDWOeserBTudzynskiB 2008 Cross-species hybridization with Fusarium verticillioides microarrays reveals new insights into Fusarium fujikuroi nitrogen regulation and the role of AreA and NMR. Eukaryotic Cell 7 1831 1846
35. TeichertSWottawaMSchönigBTudzynskiB 2006 Role of the Fusarium fujikuroi TOR kinase in nitrogen regulation and secondary metabolism. Eukaryotic Cell 5 1807 1819
36. TudzynskiB 1999 Biosynthesis of gibberellins in Gibberella fujikuroi: biomolecular aspects. Appl Microbiol Biotechnol 52 298 310
37. López-BergesMSRisapailNPrados-RosalesRCDi PietroA 2010 A nitrogen response pathway regulates virulence functions in Fusarium oxysporum via the protein kinase TOR and the bZIP protein MeaB. The Plant Cell 22 2459 2475
38. ChiangTYMarzlufGA 1994 DNA recognition by NIT2 nitrogen regulatory protein: importance of number, spacing, and orientation of GATA core elements and their flanking sequences upon NIT2 binding. Biochemistry 33 576 582
39. CunninghamTSCooperTG 1993 The Saccaromyces cerevisiae DAL80 represssor protein binds to multiple copies of GATAA-containing sequences (URSGATA). J Bacteriol 175 5851 5861
40. FrandsenFJNKristensenMBAnderssonJAGieseH 2008 Efficient four fragment cloning for the construction of vectors for targeted gene replacement in filamentous fungi. BMC Mol Biol 9 70
41. MillerJDBlackwellBA 1986 Biosynthesis of 3-acetyldeoxynivalenol and other metabolites by Fusarium culmorum HLX 1503 in a stirred jar fermenter. Can J Bot 64 1 5
42. SalehAAlvarez-VenegasRAvramovaZ 2008 An efficient chromatin immunoprecipitation (ChIP) protocol for studying histone modifications in Arabidopsis plants. Nat Protocols 3 1018 1025
43. LiHHomerN 2010 A survey of sequence alignment algorithms for next-generation sequencing. Briefings in Bioinformatics 11 473 483
44. KrawitzPRödelspergerCJägerMJostinsLBauerS 2010 Microindel detection in short-read sequence data. Bioinformatics 26 722 729
Štítky
Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
PLOS Pathogens
2011 Číslo 9
- 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
- Hostile Takeover by : Reorganization of Parasite and Host Cell Membranes during Liver Stage Egress
- HTLV-1 Propels Thymic Human T Cell Development in “Human Immune System” Rag2 gamma c Mice
- Exploiting and Subverting Tor Signaling in the Pathogenesis of Fungi, Parasites, and Viruses
- A Viral Ubiquitin Ligase Has Substrate Preferential SUMO Targeted Ubiquitin Ligase Activity that Counteracts Intrinsic Antiviral Defence