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Characterization of theugatA gene ofUstilago maydis, isolated by homology to thegatA gene ofAspergillus nidulans

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Abstract

A gene encoding a putative GABA aminotransferase (ugatA) was isolated from the basidiomyceteUstilago maydis via heterologous hybridization to the GABA aminotransferase gene (gatA) ofAspergillus nidulans. The derived amino-acid sequence ofugatA shows strong identity throughout the protein to the GABA aminotransferase enzymes fromA. nidulans andSaccharomyces cerevisiae. Northern analysis inU. maydis indicated that theugatA transcript is inducible by the ω-amino acids GABA and β-alanine, and is not subject to nitrogen catabolite repression. With the use ofugatA promoter-lacZ fusion constructs, it was demonstrated that the removal of sequences located approximately 250 by 5′ to the translational start site ofugatA (including multiple copies of a 7-bp direct repeat) resulted in the loss of induction by ω-amino acids. While theugatA gene under the control of theA. nidulans gatA promoter was able to fully complement agatA − phenotype inA. nidulans, the full-lengthugatA gene was not, suggesting a lack of expression from theU. maydis promoter inA. nidulans. AU. maydis strain with a gene disruption at theugatA locus showed decreased growth on β-alanine as a sole nitrogen source, but was able to grow on GABA as a sole nitrogen source, indicating an alternative pathway for the utilization of GABA inU. maydis.

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References

  • Alic M, Clark EK, Kornegay JR, Gold MH (1990) Transformation ofPhanerochaete chrysosporium andNeurospora crassa with adenine biosynthetic genes fromSchizophyllum commune Curr Genet 17:305–311

    Google Scholar 

  • Altschul SF, Gish W, Miller W, Myers EW, Lipman D (1990) Basic local alignment search tool. J Mol Biol 215:403–410

    Google Scholar 

  • André B, Jauniaux J-C (1990) Nucleotide sequence of the yeastUGA1 gene encoding GABA transaminase. Nucleic Acids Res 18:3049

    Google Scholar 

  • Andrianopoulos A, Hynes MJ (1988) Cloning and analysis of the positively acting regulatory geneamdR fromAspergillus nidulans. Mol Cell Biol 8:3532–3541

    Google Scholar 

  • Arndt K, Fink GR (1986) GCN4 protein, a positive transcription factor in yeast, binds general control promoters at all 5′ TGACTC 3′ sequences. Proc Natl Acad Sci USA 83:8516–8520

    Google Scholar 

  • Arst HN Jr (1976) Integrator gene inAspergillus nidulans. Nature 262:231–234

    Google Scholar 

  • Arst HN Jr, Penfold HA, Bailey CR (1978) Lactam utilization inAspergillus nidulans: evidence for a fourth gene under the control of the integrator geneintA. Mol Gen Genet 166:321–327

    Google Scholar 

  • Bailey CR, Penfold HA, Arst HN Jr (1979)cis-dominant regulatory mutations affecting the expression of GABA permease inAspergillus nidulans. Mol Gen Genet 169:79–83

    Google Scholar 

  • Banks GR, Shelton PA, Kanuga N, Holden DW, Spanos A (1993) TheUstilago maydis nar1 gene encoding nitrate reductase activity; sequence and transcriptional regulation. Gene 131:69–78

    Google Scholar 

  • Banks GR, Taylor SY (1988) Cloning of thePYR3 gene ofUstilago maydis and its use in DNA transformation. Mol Cell Biol 8:5417–5424

    Google Scholar 

  • Banuett F (1992)Ustilago maydis, the delightful blight. Trends Genet 8:174–180

    Google Scholar 

  • Bauchwitz R, Holloman WK (1990) Isolation of theREC2 gene controlling recombination inUstilago maydis. Gene 96:285–288

    Google Scholar 

  • Bölker M, Urban M, Kahmann R (1992) The a mating-type locus ofU. maydis specifies cell signalling components. Cell 68:441–450

    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

    Google Scholar 

  • Casselton LA, de la Fuente Herce A (1989) Heterologous gene expression in the basidiomycete fungusCoprinus cinereus. Curr Genet 16:35–40

    Google Scholar 

  • Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159

    Google Scholar 

  • Clutterbuck AJ (1984) Loci and linkage map of the filamentous fungusAspergillus nidulans. In: O'Brien SJ (ed) Genetic maps, vol.3. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 265–273

    Google Scholar 

  • Cove DJ (1966) The induction and repression of nitrate reductase in the fungusAspergillus nidulans. Biochem Biophys Acta 133:51–56

    Google Scholar 

  • Cunningham TS, Cooper TG (1991) Expression of theDAL80 gene, whose product is homologous to the GATA factors and is a negative regulator of multiple nitrogen catabolic genes inSaccharomyces cerevisae, is sensitive to nitrogen catabolite repression. Mol Cell Biol 11:6205–6215

    Google Scholar 

  • Daugherty JR, Rai R, El Berry H, Cooper TG (1993) Regulatory circuit for responses of nitrogen catabolic gene expression to theGLN3 andDAL80 proteins and nitrogen catabolite repression inSaccharomyces cerevisae. J Bacteriol 175:64–73

    Google Scholar 

  • Davis MA, Cobbett CS, Hynes MJ (1988) AnamdS-lacZ fusion for studying gene regulation inAspergillus. Gene 63:199–212

    Google Scholar 

  • De Biase D, Maras B, Bossa F, Barra D, John RA (1992) Protein structure of pig liver 4-aminobutyrate aminotransferase and comparison with a cDNA-deduced sequence. Eur J Biochem 208:351–357

    Google Scholar 

  • Ehinger A, Denison SH, May GS (1990) Sequence, organization and expression of the core histone genes ofAspergillus nidulans. Mol Gen Genet 222:416–424

    Google Scholar 

  • Elder RT, Lou EY, Davis RW (1983) RNA from the yeast transposable element Tyl has both ends in the direct repeats, a structure similar to retrovirus RNA. Proc Natl Acad Sci USA 80:2432–2436

    Google Scholar 

  • Falco SC, Dumas KS, Livak KJ (1985) Nucleotide sequence of the yeastILV2 gene which encodes acetolactate synthase. Nucleic Acids Res 13:4011–4027

    Google Scholar 

  • Fisher HF (1985) Glutamate, glutamine, glutathione and related compounds. Methods Enzymol 113:16–27

    Google Scholar 

  • Fotheringham S, Holloman WK (1990) Pathways of transformation inUstilago maydis determined by DNA conformation. Genetics 124:833–843

    Google Scholar 

  • Fotheringham S, Holloman WK (1991) Extrachromosomal recombination is deranged in therec2 mutant ofUstilago maydis. Genetics 129:1053–1060

    Google Scholar 

  • Froeliger EH, Leong SA (1991) The a mating-type ofUstilago maydis are idiomorphs. Gene 100:113–122

    Google Scholar 

  • Grenson M, Muyldermans F, Broman K, Vissers S (1987) 4-amino-butyric acid (GABA) uptake in Baker's yeastSaccharomyces cerevisiae is mediated by the general amino-acid permease, the proline permease and a GABA-specific permease integrated into the GABA-catabolic pathway. Life Sci Adv 6:35–39

    Google Scholar 

  • Higgins DG, Bleasby AJ, Fuchs R (1992) Clustal V: improved software for multiple sequence alignment. CABIOS 8:189–191

    Google Scholar 

  • Holliday R (1974)Ustilago maydis. In: King RC (ed) Handbook of genetics. Plenum, New York pp 575–595

    Google Scholar 

  • Hynes MJ, Pateman JA (1970) The genetic analysis of regulation of amidase synthesis inAspergillus nidulans. II: Mutants able to utilize acrylamide. Mol Cell Genet 108:97–106

    Google Scholar 

  • Katz ME, Hynes MJ (1989) Characterization of theamdR-control-ledlamA andlamB genes ofAspergillus nidulans. Generics 122:311–339

    Google Scholar 

  • Keon JPR, James CS, Court S, Baden-Daintree C, Bailey AM, Burden RS, Bard M, Hargreaves JA (1994) Isolation of theERG2 gene, encoding sterol8 →7isomerase, from the rice blast fungusMagnaporthe grisea and its expression in the maize smut pathogenUstilago maydis. Curr Genet 25:531–537

    Google Scholar 

  • Kozak M (1984) Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. Nucleic Acids Res 12:857–872

    Google Scholar 

  • Kozak M (1987) An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res 15:8125–8148

    Google Scholar 

  • Kwon O-S, Park J, Churchich JE (1992) Brain 4-aminobutyrate aminotransferase: isolation and sequence of a cDNA encoding the enzyme. J Biol Chem 267:7215–7216

    Google Scholar 

  • Lee SB, Taylor JW (1989) Isolation of DNA from fungal mycelia and single spores. In: Innis MA, Gelford DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic Press Inc., Harcourt Brace Jovanovich, USA, pp 282–287

    Google Scholar 

  • Lewis CM, Fincham JRS (1970 a) Genetics of nitrate reductase inUstilago maydis. Genet Res 16:151–63

    Google Scholar 

  • Lewis CM, Fincham JRS (1970a) Regulation of nitrate reductase in the basidiomyceteUstilago maydis. J Bacteriol 103:55–61

    Google Scholar 

  • Marck C (1988) ‘DNA Strider’: a ‘C’ program for the fast analysis of DNA and protein sequences on the Apple Macintosh family of computers. Nucleic Acids Res 16:1829–1836

    Google Scholar 

  • Oakley CE, Kretz PL, Oakley BR (1987) Cloning of theriboB2 locus ofAspergillus nidulans. Gene 53:293–298

    Google Scholar 

  • Pearson WR, Lipman DJ (1988) Improved tools for biological sequence analysis. Proc Natl Acad Sci USA 85:2444–2448

    Google Scholar 

  • Prestridge DS (1991) SIGNAL SCAN: a computer program that scans DNA sequences for eucaryotic transcriptional elements. CABIOS 7:203–206

    Google Scholar 

  • Ramos F, El Guezzar M, Grenson M, Wiame J-M (1985) Mutations affecting the enzymes involved in the utilization of 4-amino-butyric acid as nitrogen source by the yeastSaccharomyces cerevisae. Eur J Biochem 149:401–404

    Google Scholar 

  • Richardson IB, Hurley SK, Hynes MJ (1989) Cloning and molecular characterization of theamdR controlledgatA gene ofAspergillus nidulans. Mol Cell Genet 217:118–125

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual (2nd edn.) Cold Spring Harbor Laboratory Cold Spring Harbor, New York

    Google Scholar 

  • Spanos A, Kanuga N, Holden DW, Banks GR (1992) TheUstilago maydis pyr3 gene: sequence and transcriptional analysis. Gene 117:73–79

    Google Scholar 

  • Talibi D, Grenson M, André B (1995)Cis- andtrans-acting elements determining induction of the genes of the γ-aminobutyrate (GABA) utilization pathway inSaccharomyces cerevisiae. Nucleic Acids Res 23:550–557

    Google Scholar 

  • Thatcher TH, Gorovosky MA (1994) Phylogenetic analysis of the core histories H2A, H2B, H3 and H4. Nucleic Acid Res 22 174–179

    Google Scholar 

  • Tsukuda T, Carleton S, Fotheringham S, Holloman WK (1988) Isolation and characterization of an autonomously replicating sequence fromUstilago maydis. Mol Cell Biol 8:3703–3709

    Google Scholar 

  • Tsukuda T, Bauchwitz R, Holloman WK (1989) Isolation of theREC1 gene controlling recombination inUstilago maydis. Gene 85:335–341

    Google Scholar 

  • Van Gorcom REM, Punt PJ, Pouwels PH, van den Hondel CAMJJ (1986) A system for the analysis of expression signals inAspergillus. Gene 48:211–217

    Google Scholar 

  • Vissers S, André B, Muyldermans F, Grenson M (1989) Positive and negative regulatory elements control the expression of theUGA4 gene coding for the inducible 4-aminobutyic-acid-specific permease inSaccharomyces cerevisiae. Eur J Biochem 181:357–361

    Google Scholar 

  • Vissers S, André B, Muyldermans F, Grenson M (1990) Induction of 4-aminobutyrate and urea-catabolic pathways inSaccharomyces cerevisiae. Eur J Biochem 187:611–616

    Google Scholar 

  • Wahle E, Keller W (1992) The biochemistry of 3′-end cleavage and polyadenylation of messenger RNA precursors. Annu Rev Biochem 61:419–440

    Google Scholar 

  • Wang J, Holden DW, Leong SA (1988) Gene transfer system for the phytopathogenic fungusUstilago maydis. Proc Natl Acad Sci USA 85:865–869

    Google Scholar 

  • Wray JL, Kinghorn JR (eds) (1989) Molecular and genetic aspects of nitrate assimilation Oxford University Press, UK

    Google Scholar 

  • Yonaha K, Nishie M, Minei H, Toyama S (1983) Distribution of ω-amino acid: pyruvate transaminase and aminobutyate: α-ketoglutarate transaminase in microorganisms. Agric Biol Chem 47:2257–2265

    Google Scholar 

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Authors and Affiliations

  1. Department of Genetics, The University of Melbourne, Parkville, 3052, Victoria, Australia

    Melissa J. Straffon, Michael J. Hynes & Meryl A. Davis

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  1. Melissa J. Straffon
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  2. Michael J. Hynes
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  3. Meryl A. Davis
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Communicated by O.C. Yoder

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Straffon, M.J., Hynes, M.J. & Davis, M.A. Characterization of theugatA gene ofUstilago maydis, isolated by homology to thegatA gene ofAspergillus nidulans . Curr Genet 29, 360–369 (1996). https://doi.org/10.1007/BF02208617

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  • DOI: https://doi.org/10.1007/BF02208617

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