Secondary Metabolism and Development Is Mediated by LlmF Control of VeA Subcellular Localization in
Secondary metabolism and development are linked in Aspergillus through the conserved regulatory velvet complex composed of VeA, VelB, and LaeA. The founding member of the velvet complex, VeA, shuttles between the cytoplasm and nucleus in response to alterations in light. Here we describe a new interaction partner of VeA identified through a reverse genetics screen looking for LaeA-like methyltransferases in Aspergillus nidulans. One of the putative LaeA-like methyltransferases identified, LlmF, is a negative regulator of sterigmatocystin production and sexual development. LlmF interacts directly with VeA and the repressive function of LlmF is mediated by influencing the localization of VeA, as over-expression of llmF decreases the nuclear to cytoplasmic ratio of VeA while deletion of llmF results in an increased nuclear accumulation of VeA. We show that the methyltransferase domain of LlmF is required for function; however, LlmF does not directly methylate VeA in vitro. This study identifies a new interaction partner for VeA and highlights the importance of cellular compartmentalization of VeA for regulation of development and secondary metabolism.
Vyšlo v časopise:
Secondary Metabolism and Development Is Mediated by LlmF Control of VeA Subcellular Localization in. PLoS Genet 9(1): e32767. doi:10.1371/journal.pgen.1003193
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1003193
Souhrn
Secondary metabolism and development are linked in Aspergillus through the conserved regulatory velvet complex composed of VeA, VelB, and LaeA. The founding member of the velvet complex, VeA, shuttles between the cytoplasm and nucleus in response to alterations in light. Here we describe a new interaction partner of VeA identified through a reverse genetics screen looking for LaeA-like methyltransferases in Aspergillus nidulans. One of the putative LaeA-like methyltransferases identified, LlmF, is a negative regulator of sterigmatocystin production and sexual development. LlmF interacts directly with VeA and the repressive function of LlmF is mediated by influencing the localization of VeA, as over-expression of llmF decreases the nuclear to cytoplasmic ratio of VeA while deletion of llmF results in an increased nuclear accumulation of VeA. We show that the methyltransferase domain of LlmF is required for function; however, LlmF does not directly methylate VeA in vitro. This study identifies a new interaction partner for VeA and highlights the importance of cellular compartmentalization of VeA for regulation of development and secondary metabolism.
Zdroje
1. BrownDW, YuJH, KelkarHS, FernandesM, NesbittTC, et al. (1996) Twenty-five coregulated transcripts define a sterigmatocystin gene cluster in Aspergillus nidulans. Proc Natl Acad Sci U S A 93: 1418–1422.
2. BokJW, KellerNP (2004) LaeA, a regulator of secondary metabolism in Aspergillus spp. Eukaryot Cell 3: 527–535.
3. BayramO, KrappmannS, NiM, BokJW, HelmstaedtK, et al. (2008) VelB/VeA/LaeA complex coordinates light signal with fungal development and secondary metabolism. Science 320: 1504–1506.
4. Sarikaya BayramO, BayramO, ValeriusO, ParkHS, IrnigerS, et al. (2010) LaeA control of velvet family regulatory proteins for light-dependent development and fungal cell-type specificity. PLoS Genet 6: e1001226 doi:10.1371/journal.pgen.1001226.
5. BayramO, BrausGH (2012) Coordination of secondary metabolism and development in fungi: the velvet family of regulatory proteins. FEMS Microbiol Rev 36: 1–24.
6. SuguiJA, PardoJ, ChangYC, MüllbacherA, ZaremberKA, et al. (2007) Role of laeA in the regulation of alb1, gliP, conidial morphology, and virulence in Aspergillus fumigatus. Eukaryot Cell 6: 1552–1561.
7. BokJW, BalajeeSA, MarrKA, AndesD, NielsenKF, et al. (2005) LaeA, a regulator of morphogenetic fungal virulence factors. Eukaryot Cell 4: 1574–1582.
8. MyungK, ZitomerNC, DuvallM, GlennAE, RileyRT, et al. (2012) The conserved global regulator VeA is necessary for symptom production and mycotoxin synthesis in maize seedlings by Fusarium verticillioides. Plant Pathology 61: 152–160.
9. WiemannP, BrownDW, KleigreweK, BokJW, KellerNP, et al. (2010) FfVel1 and FfLae1, components of a velvet-like complex in Fusarium fujikuroi, affect differentiation, secondary metabolism and virulence. Mol Microbiol 77: 972–994.
10. WuD, OideS, ZhangN, ChoiMY, TurgeonBG (2012) ChLae1 and ChVel1 regulate T-toxin production, virulence, oxidative stress response, and development of the maize pathogen Cochliobolus heterostrophus. PLoS Pathog 8: e1002542 doi:10.1371/journal.ppat.1002542.
11. AmaikeS, KellerNP (2009) Distinct roles for VeA and LaeA in development and pathogenesis of Aspergillus flavus. Eukaryot Cell 8: 1051–1060.
12. DuranRM, CaryJW, CalvoAM (2009) The role of veA on Aspergillus flavus infection of peanuts, corn and cotton. Open Mycology Journal 3: 27–36.
13. JiangJ, LiuX, YinY, MaZ (2011) Involvement of a velvet protein FgVeA in the regulation of asexual development, lipid and secondary metabolisms and virulence in Fusarium graminearum. PLoS ONE 6: e28291 doi:10.1371/journal.pone.0028291.
14. MerhejJ, UrbanM, Hammond-KosackKE, Richard-ForgetF, BarreauC (2011) The velvet gene, FgVe1, affects fungal development and positively regulates trichothecene biosynthesis and pathogenicity in Fusarium graminearum. Mol Plant Pathol doi:10.1111/j.1364-3703.2011.00755.x.
15. JiangJ, YunY, LiuY, MaZ (2012) FgVELB is associated with vegetative differentiation, secondary metabolism and virulence in Fusarium graminearum. Fungal Genet Biol 49: 653–662.
16. LeeJ, MyongK, KimJ-E, KimH-K, YunS-H, et al. (2012) FgVelB globally regulates sexual reproduction, mycotoxin production and pathogenicity in the cereal pathogen Fusarium graminearum. Microbiology 158: 1723–1733.
17. BokJW, NoordermeerD, KaleSP, KellerNP (2006) Secondary metabolic gene cluster silencing in Aspergillus nidulans. Mol Microbiol 61: 1636–1645.
18. StinnettSM, EspesoEA, CobenoL, Araujo-BazanL, CalvoAM (2007) Aspergillus nidulans VeA subcellular localization is dependent on the importin alpha carrier and on light. Mol Microbiol 63: 242–255.
19. KimH, HanK, KimK, HanD, JahngK, et al. (2002) The veA gene activates sexual development in Aspergillus nidulans. Fungal Genet Biol 37: 72–80.
20. PurschwitzJ, MüllerS, KastnerC, SchöserM, HaasH, et al. (2008) Functional and physical interaction of blue- and red-light sensors in Aspergillus nidulans. Curr Biol 18: 255–259.
21. PurschwitzJ, MüllerS, FischerR (2009) Mapping the interaction sites of Aspergillus nidulans phytochrome FphA with the global regulator VeA and the White Collar protein LreB. Mol Genet Genomics 281: 35–42.
22. NiM, YuJH (2007) A novel regulator couples sporogenesis and trehalose biogenesis in Aspergillus nidulans. PLoS ONE 2: e970 doi:10.1371/journal.pone.0000970.
23. CalvoAM (2008) The VeA regulatory system and its role in morphological and chemical development in fungi. Fungal Genet Biol 45: 1053–1061.
24. PalmerJM, MallaredyS, PerryDW, SanchezJF, TheisenJM, et al. (2010) Telomere position effect is regulated by heterochromatin-associated proteins and NkuA in Aspergillus nidulans. Microbiology 156: 3522–3531.
25. EdgarRC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32: 1792–1797.
26. KaganRM, ClarkeS (1994) Widespread occurrence of three sequence motifs in diverse S-adenosylmethionine-dependent methyltransferases suggests a common structure for these enzymes. Arch Biochem Biophys 310: 417–427.
27. PetrossianTC, ClarkeSG (2009) Multiple motif scanning to identify methyltransferases from the yeast proteome. Mol Cell Proteomics 8: 1516–1526.
28. ShaabanMI, BokJW, LauerC, KellerNP (2010) Suppressor mutagenesis identifies a velvet complex remediator of Aspergillus nidulans secondary metabolism. Eukaryot Cell 9: 1816–1824.
29. Araújo-BazanL, DhingraS, ChuJ, Fernández-MartínezJ, CalvoAM, et al. (2009) Importin alpha is an essential nuclear import carrier adaptor required for proper sexual and asexual development and secondary metabolism in Aspergillus nidulans. Fungal Genet Biol 46: 506–515.
30. TrojerP, DanglM, BauerI, GraessleS, LoidlP, et al. (2004) Histone methyltransferases in Aspergillus nidulans: evidence for a novel enzyme with a unique substrate specificity. Biochemistry (Mosc) 43: 10834–10843.
31. NardozziJD, LottK, CingolaniG (2010) Phosphorylation meets nuclear import: a review. Cell Commun Signal 8: 32.
32. WhitmarshAJ, DavisRJ (2000) Regulation of transcription factor function by phosphorylation. Cell Mol Life Sci 57: 1172–1183.
33. BernreiterA, RamónA, Fernández-MartínezJ, BergerH, Araújo-BazanL, et al. (2007) Nuclear export of the transcription factor NirA is a regulatory checkpoint for nitrate induction in Aspergillus nidulans. Mol Cell Biol 27: 791–802.
34. BergerH, PachlingerR, MorozovI, GollerS, NarendjaF, et al. (2006) The GATA factor AreA regulates localization and in vivo binding site occupancy of the nitrate activator NirA. Mol Microbiol 59: 433–446.
35. BeckT, HallMN (1999) The TOR signalling pathway controls nuclear localization of nutrient-regulated transcription factors. Nature 402: 689–692.
36. PelegY, AddisonR, AramayoR, MetzenbergRL (1996) Translocation of Neurospora crassa transcription factor NUC-1 into the nucleus is induced by phosphorus limitation. Fungal Genet Biol 20: 185–191.
37. Maggio-HallLA, KellerNP (2004) Mitochondrial beta-oxidation in Aspergillus nidulans. Mol Microbiol 54: 1173–1185.
38. ChandaA, RozeLV, KangS, ArtymovichKA, HicksGR, et al. (2009) A key role for vesicles in fungal secondary metabolism. Proc Natl Acad Sci U S A 106: 19533–19538.
39. HynesMJ, MurraySL, KhewGS, DavisMA (2008) Genetic analysis of the role of peroxisomes in the utilization of acetate and fatty acids in Aspergillus nidulans. Genetics 178: 1355–1369.
40. BlumensteinA, VienkenK, TaslerR, PurschwitzJ, VeithD, et al. (2005) The Aspergillus nidulans phytochrome FphA represses sexual development in red light. Curr Biol 15: 1833–1838.
41. ShimizuK, HicksJK, HuangTP, KellerNP (2003) Pka, Ras and RGS protein interactions regulate activity of AflR, a Zn(II)2Cys6 transcription factor in Aspergillus nidulans. Genetics 165: 1095–1104.
42. BayramO, BayramÖS, AhmedYL, MaruyamaJ-I, ValeriusO, et al. (2012) The Aspergillus nidulans MAPK module AnSte11-Ste50-Ste7-Fus3 controls development and secondary metabolism. PLoS Genet 8: e1002816 doi:10.1371/journal.pgen.1002816.
43. SmithWA, SchurterBT, Wong-StaalF, DavidM (2004) Arginine methylation of RNA helicase a determines its subcellular localization. J Biol Chem 279: 22795–22798.
44. ShimizuK, KellerNP (2001) Genetic involvement of a cAMP-dependent protein kinase in a G protein signaling pathway regulating morphological and chemical transitions in Aspergillus nidulans. Genetics 157: 591–600.
45. Sambrook J, Russell DW (2001) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.
46. ArnaudMB, ChibucosMC, CostanzoMC, CrabtreeJ, InglisDO, et al. (2010) The Aspergillus Genome Database, a curated comparative genomics resource for gene, protein and sequence information for the Aspergillus research community. Nucleic Acids Res 38: D420–427.
47. YuJH, HamariZ, HanKH, SeoJA, Reyes-DomínguezY, et al. (2004) Double-joint PCR: a PCR-based molecular tool for gene manipulations in filamentous fungi. Fungal Genet Biol 41: 973–981.
48. YangL, UkilL, OsmaniA, NahmF, DaviesJ, et al. (2004) Rapid production of gene replacement constructs and generation of a green fluorescent protein-tagged centromeric marker in Aspergillus nidulans. Eukaryot Cell 3: 1359–1362.
49. SzewczykE, NayakT, OakleyCE, EdgertonH, XiongY, et al. (2006) Fusion PCR and gene targeting in Aspergillus nidulans. Nat Protoc 1: 3111–3120.
50. ShaabanM, PalmerJM, El-NaggarWA, El-SokkaryMA, HabibE-SE, et al. (2010) Involvement of transposon-like elements in penicillin gene cluster regulation. Fungal Genet Biol 47: 423–432.
51. MillerBL, MillerKY, TimberlakeWE (1985) Direct and indirect gene replacements in Aspergillus nidulans. Mol Cell Biol 5: 1714–1721.
52. TsitsigiannisDI, ZarnowskiR, KellerNP (2004) The lipid body protein, PpoA, coordinates sexual and asexual sporulation in Aspergillus nidulans. J Biol Chem 279: 11344–11353.
53. van den EntF, LöweJ (2006) RF cloning: a restriction-free method for inserting target genes into plasmids. J Biochem Biophys Methods 67: 67–74.
54. BuschS, SchwierEU, NahlikK, BayramO, HelmstaedtK, et al. (2007) An eight-subunit COP9 signalosome with an intact JAMM motif is required for fungal fruit body formation. Proc Natl Acad Sci U S A 104: 8089–8094.
55. ChoJ, YunS, JangY, ChaM, KwonN, et al. (2003) Identification and cloning of jipA encoding a polypeptide that interacts with a homolog of yeast Rad6, UVSJ in Aspergillus nidulans. J Microbiol 41: 46–51.
56. VojtekAB, HollenbergSM, CooperJA (1993) Mammalian Ras interacts directly with the serine/threonine kinase Raf. Cell 74: 205–214.
57. GietzRD, SchiestlRH (2007) Frozen competent yeast cells that can be transformed with high efficiency using the LiAc/SS carrier DNA/PEG method. Nat Protoc 2: 1–4.
58. DongH, RenS, ZhangB, ZhouY, Puig-BasagoitiF, et al. (2008) West Nile virus methyltransferase catalyzes two methylations of the viral RNA cap through a substrate-repositioning mechanism. J Virol 82: 4295–4307.
59. SchäggerH (2006) Tricine-SDS-PAGE. Nat Protoc 1: 16–22.
60. Momany M (2001) Cell biology of the duplication cycle in fungi. In: Talbot N, editor. Molecular and cellular biology of filamentous fungi: a practical approach: Oxford University Press, USA. pp. 119–124.
61. KatzJE, DlakićM, ClarkeS (2003) Automated identification of putative methyltransferases from genomic open reading frames. Mol Cell Proteomics 2: 525–540.
Štítky
Genetika Reprodukčná medicínaČlánok vyšiel v časopise
PLOS Genetics
2013 Číslo 1
- Je „freeze-all“ pro všechny? Odborníci na fertilitu diskutovali na virtuálním summitu
- Gynekologové a odborníci na reprodukční medicínu se sejdou na prvním virtuálním summitu
Najčítanejšie v tomto čísle
- Function and Regulation of , a Gene Implicated in Autism and Human Evolution
- Comprehensive Methylome Characterization of and at Single-Base Resolution
- Susceptibility Loci Associated with Specific and Shared Subtypes of Lymphoid Malignancies
- An Insertion in 5′ Flanking Region of Causes Blue Eggshell in the Chicken