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Promoter analysis of the avirulence gene Avr9 of the fungal tomato pathogen Cladosporium fulvum in the model filamentous fungus Aspergillus nidulans

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Abstract.

The promoter of avirulence gene Avr9 of the fungal tomato pathogen Cladosporium fulvum contains 12 sequences within a region of 0.6 kb that are reminiscent of the binding sequences of the GATA-type regulator involved in nitrogen utilisation of the filamentous fungi Aspergillus nidulans and Neurospora crassa. Mutational analysis of this 0.6-kb promoter region, fused to the β-glucuronidase reporter gene, revealed that two regions, each containing two TAGATA boxes in inverted orientation and overlapping by two base pairs, are important for induction of Avr9 promoter activity in A. nidulans. Each overlapping TAGATA box differentially affected Avr9 promoter activity when shifted apart by nucleotide insertions. The other regions, which do not contain two overlapping TAGATA boxes have no, or only a limited, contribution to the inducibility of promoter activity.

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References

  • Beckett D (2001) Regulated assembly of transcription factors and control of transcription initiation. J Mol Biol 314:335–352

    Google Scholar 

  • Bradford MM (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

    Article  CAS  PubMed  Google Scholar 

  • Chiang T, Marzluf GA (1995) Binding affinity and functional significance of NIT2 and NIT4 binding sites in the promoter of the highly regulated nit-3 gene, which encodes nitrate reductase in Neurospora crassa. J Bacteriol 177:6093–6099

    CAS  PubMed  Google Scholar 

  • Chiang T, Rai R, Cooper TG, Marzluf GA (1994) DNA binding site specificity of the Neurospora global nitrogen protein NIT2: analysis with mutated binding sites. Mol Gen Genet 245:512–516

    CAS  PubMed  Google Scholar 

  • Christensen T, Hynes MJ, Davis MA (1998) Role of the regulatory gene areA of Aspergillus oryzae in nitrogen metabolism. Appl Environ Microbiol 64:3232–3237

    CAS  PubMed  Google Scholar 

  • Davis MA, Hynes MJ (1987) Complementation of areA-regulatory gene mutations of Aspergillus nidulans by the heterologous regulatory gene nit-2 of Neurospora crassa. Proc Natl Acad Sci USA 84:3753–3757

    CAS  PubMed  Google Scholar 

  • De Wit PJGM (1995) Fungal avirulence genes and plant resistance genes: unraveling the molecular basis of gene-for-gene interactions. Adv Bot Res 21:147–185

    Google Scholar 

  • Froeliger E, Carpenter B (1996) NUT1, a major nitrogen regulatory gene in Magnaporthe grisea, is dispensable for pathogenicity. Mol Gen Genet 25:647–656

    Article  Google Scholar 

  • Fu YH, Marzluf GA (1990) nit-2, the major positive-acting nitrogen regulatory gene of Neurospora crassa, encodes a sequence-specific DNA-binding protein. Proc Natl Acad Sci USA 87:5331–5335

    CAS  PubMed  Google Scholar 

  • Haas HB, Marzluf GA (1995) NRE, the major nitrogen regulatory protein of Penicillium chrysogenum, binds specifically to elements in the intergenic promoter regions of nitrate assimilation and penicillin biosynthetic gene clusters. Curr Genet 28:177–183

    CAS  PubMed  Google Scholar 

  • Hawkins AR, Lamb HK, Radford A, Moore JD (1994) Evolution of transcription-regulating proteins by enzyme recruitment: molecular models for nitrogen metabolite repression and ethanol utilisation in eukaryotes. Gene 146:145–158

    CAS  PubMed  Google Scholar 

  • Hensel M, Tang CM, Arst HN, Holden DW (1995) Regulation of fungal extracellular proteases and their role in mammalian pathogenesis. Can J Bot 73 [Suppl 1]:S1065–S1070

    Google Scholar 

  • Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusion: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907

    CAS  PubMed  Google Scholar 

  • Johnstone LL, McCabe PC, Greaves P, Gurr SJ, Cole GE, Brow MAD, Unkles SE, Clutterbuck AJ, Kinghorn JR, Innes MA (1990) Isolation and characterisation of the crnA-niiA-niaD gene cluster for nitrate assimilation in Aspergillus nidulans. Gene 90:181–192

    CAS  PubMed  Google Scholar 

  • Joosten MHAJ, De Wit PJGM (1999) The tomato–Cladosporium fulvum interaction: a versatile experimental system to study plant–pathogen interactions. Annu Rev Phytopathol 37:335–367

    CAS  PubMed  Google Scholar 

  • Marzluf GA (1997) Genetic regulation of nitrogen metabolism in the fungi. Microbiol Mol Biol Rev 61:17–32

    CAS  PubMed  Google Scholar 

  • McCabe AP, Vanhanen S, Gelpke MDS, Van de Vondervoort PJI, Arst HN, Visser J (1998) Identification, cloning and sequence of the Aspergillus niger areA wide domain regulatory gene controlling nitrogen utilisation. Biochem Biophys Acta 1396:163–168

    Google Scholar 

  • Muro-Pastor MI, Gonzalez R, Strauss J, Narendja F, Scazzocchio C (1999) The GATA factor AreA is essential for chromatin remodelling in a eukaryotic bidirectional promoter. EMBO J 18:1584–1597

    CAS  PubMed  Google Scholar 

  • Narendja F, Goller SP, Wolscheck M, Strauss J (2002) Nitrate and the GATA factor AreA are necessary for in vivo binding of NirA, the pathway-specific transcriptional activator of Aspergillus nidulans. Mol Microbiol 44:573–583

    Article  CAS  PubMed  Google Scholar 

  • Pérez-García A, Snoeijers SS, Joosten MHAJ, Goosen T, De Wit PJGM (2001) Expression of the avirulence gene Avr9 of the fungal tomato pathogen Cladosporium fulvum is regulated by the global nitrogen response factor NRF1. Mol Plant-Microbe Interact 14:316–325

    Google Scholar 

  • Punt PJ, Strauss J, Smit R, Kinghorn JR, Van den Hondel CAMJJ, Scazzocchio C (1995) The intergenic region between divergently transcribed niiA and niaD genes of Aspergillus nidulans contains multiple NirA binding sites which act bidirectionally. Mol Cell Biol 15:5688–5699

    CAS  PubMed  Google Scholar 

  • Ravagnani A, Gorfinkiel L, Langdon T, Diallinas G, Adjadj E, Demais S, Gorton D, Arst HN, Scazzocchio C (1997) Subtle hydrophobic interactions between the seventh residue of the zinc finger loop at the first base of an HGATAR sequence determine promoter-specific recognition by the Aspergillus nidulans GATA factor AreA. EMBO J 16:3974–3986

    CAS  PubMed  Google Scholar 

  • Screen S, Bailey A, Charnley K, Cooper R, Clarkson J (1998) Isolation of a nitrogen response regulator gene (nrr1) from Metarhizium anisopliae. Gene 221:17–24

    Article  CAS  PubMed  Google Scholar 

  • Snoeijers SS (2000) Regulation of the avirulence gene Avr9 of the fungal tomato pathogen Cladosporium fulvum. PhD thesis, Wageningen University, Wageningen, pp 120

  • Snoeijers SS, Vossen P, Goosen T, Van den Broek HWJ, De Wit PJGM (1999) Transcription of the avirulence gene Avr9 of the fungal tomato pathogen Cladosporium fulvum is regulated by a GATA-type transcription factor in Aspergillus nidulans. Mol Gen Genet 261:653–659

    Article  CAS  PubMed  Google Scholar 

  • Snoeijers SS, Pérez-García A, Joosten MAHJ, De Wit PJGM (2000) The effect of nitrogen on disease development and gene expression in bacterial and fungal plant pathogens. Eur J Plant Pathol 106:493–506

    Article  CAS  Google Scholar 

  • Svaren J, Hörz W (1995) Interplay between nucleosomes and transcription factors at the yeast Pho5 promoter. Semin Cell Biol 6:177–183

    Article  CAS  PubMed  Google Scholar 

  • Thijs H, Garde J, Goosen T, Tomsett B, Swart K, Heyting C, Van den Broek HWJ (1995) Polarity of meiotic gene conversion is 5' to 3' within the niaD gene of Aspergillus nidulans. Mol Gen Genet 247:343–350

    CAS  PubMed  Google Scholar 

  • Tilburn J, Scazzocchio C, Taylor GG, Zabicky-Zissman JH, Lockington RA, Davies RW (1983) Transformation by integration in Aspergillus nidulans. Gene 26:205–221

    CAS  PubMed  Google Scholar 

  • Tudzynski B, Homann V, Feng B, Marzluf GA (1999) Isolation, characterization and disruption of the areA nitrogen regulatory gene of Gibberella fujikuroi. Mol Gen Genet 261:106–114

    CAS  PubMed  Google Scholar 

  • Van den Ackerveken GFJM, Van Kan JAL, De Wit PJGM (1992) Molecular analysis of the avirulence gene Avr9 of the fungal tomato pathogen Cladosporium fulvum fully supports the gene-for-gene hypothesis. Plant J 2:359–366

    PubMed  Google Scholar 

  • Van den Ackerveken GFJM, Dunn RM, Cozijnsen TJ, Vossen P, Van den Broek HWJ, De Wit PJGM (1994) Nitrogen limitation induces expression of the avirulence gene Avr9 in the tomato pathogen Cladosporium fulvum. Mol Gen Genet 243:277–285

    PubMed  Google Scholar 

  • Van Kan JAL, Van den Ackerveken GFJM, De Wit PJGM (1991) Cloning and characterisation of cDNA of avirulence gene Avr9 of the fungal tomato pathogen Cladosporium fulvum, causal agent of tomato leaf mold. Mol Plant-Microbe Interact 4:52–59

    Google Scholar 

  • Wernars K, Goosen T, Wennekes LM, Visser J, Bos CJ, Van den Broek HWJ, Van Gorcom RF, Van de Hondel CAMJJ, Pouwels PH (1985) Gene amplification in Aspergillus nidulans by transformation with vectors containing the amdS gene. Curr Genet 9:361–368

    CAS  PubMed  Google Scholar 

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Acknowledgements.

S.S.S. was supported by the Dutch Earth and Life Science Foundation (ALW project number 805.45.006), which is subsidised by the Netherlands Organisation for Scientific Research (NWO). A.P.-G. was supported by the EC-TMR program (ERBFMBICT 961539).

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Author notes

  1. Sandor S. Snoeijers

    Present address: Kreatech Biotechnology B.V., Vlierweg 20, 1032 LG, Amsterdam, The Netherlands

  2. Alejandro Pérez-García

    Present address: Departamento de Microbiología, Facultad de Ciencias, Universidad de Málaga, Campus Universitario de Teatinos s/n, 29071, Málaga, Spain

Authors and Affiliations

  1. Laboratory of Phytopathology, Department of Plant Sciences, Wageningen University, Binnenhaven 5, 6709 PD, Wageningen, The Netherlands

    Sandor S. Snoeijers, Alejandro Pérez-García & Pierre J. G. M. De Wit

  2. Laboratory of Genetics, Department of Biomolecular Sciences, Wageningen University, Dreijenlaan 2, 6703 HA, Wageningen, The Netherlands

    Sandor S. Snoeijers & Theo Goosen

Authors
  1. Sandor S. Snoeijers
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  2. Alejandro Pérez-García
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  3. Theo Goosen
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  4. Pierre J. G. M. De Wit
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Correspondence to Pierre J. G. M. De Wit.

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Communicated by U. Kück

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Snoeijers, S.S., Pérez-García, A., Goosen, T. et al. Promoter analysis of the avirulence gene Avr9 of the fungal tomato pathogen Cladosporium fulvum in the model filamentous fungus Aspergillus nidulans . Curr Genet 43, 96–102 (2003). https://doi.org/10.1007/s00294-003-0374-7

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  • DOI: https://doi.org/10.1007/s00294-003-0374-7

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