Skip to main content
SpringerLink
Log in

Cloning and molecular characterisation of the amdR controlled gatA gene of Aspergillus nidulans

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

The gamma-amino-n-butyrate transaminase gene (gatA) of Aspergillus nidulans is one of several genes under positive control by the regulatory gene amdR (also called intA). The gatA gene has been cloned from a cosmid library by complementation of a gatA mutation. The sequence of a 2.6 kb genomic fragment containing gatA has been determined. An open reading frame of 1497 bp within this sequences is interrupted by three putative introns and predicts a protein of 55 kDa. Northern analysis confirms control of gatA RNA levels by amdR and also indicates that gatA is not strongly regulated by areA-mediated nitrogen metabolite repression. A. nidulans transformants containing multiple copies of a plasmid carrying an 88 bp fragment from the 5′ untranscribed region of gatA grew poorly on substrates whose utilisation is dependent on genes controlled by amdR. This indicated titration of limiting amounts of the amdR gene product by this 88 bp fragment. Comparison of this sequence with the 5′ region of the coregulated gene, amdS, reveals probable sites of action for the amdR protein.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Andrianopoulos A, Hynes MJ (1988) Cloning and analysis of the positively acting regulatory gene amdR from Aspergillus 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′TGACTG3′ sequences. Proc Natl Acad Sci USA 83:8516–8520

    Google Scholar 

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

    Google Scholar 

  • Arst HN, Cove DJ (1973) Nitrogen metabolite repression in Aspergillus nidulans. Mol Gen Genet 126:111–141

    Google Scholar 

  • Arst HN, Penfold HA, Bailey CR (1978) Lactam utilisation in Aspergillus nidulans: Evidence for a fourth gene under the control of the integrator gene intA. Mol Gen Genet 166:321–327

    Google Scholar 

  • Bailey CR, Penfold HA, Arst HN (1979) Cis-dominant regulatory mutations affecting the GABA permease in Aspergillus nidulans. Mol Gen Genet 169:79–83

    Google Scholar 

  • Bailey CR, Arst HN, Penfold HA (1980) A third gene affecting GABA transaminase levels in Aspergillus nidulans. Genet Res Camb 36:167–180

    Google Scholar 

  • Baum JA, Geever R, Giles NH (1987) Expression of qa-1F activator protein: Identification of upstream binding sites in the qa gene cluster and localisation of the DNA-binding domain. Mol Cell Biol 7:1256–1266

    Google Scholar 

  • Bram RJ, Lue NF, Kornberg RD (1986) A GAL family of upstream activating sequences in yeast: Roles in both induction and repression of transcription. EMBO J 5:603–608

    Google Scholar 

  • Breathnach R, Chambon P (1981) Organization and expression of eukaryotic split genes coding for proteins. Annu Rev Biochem 50:349–383

    Google Scholar 

  • Chodosh LA, Baldwin AS, Carthew RW, Sharp PA (1988) Human CCAAT-binding proteins have heterologous subunits. Cell 53:11–24

    Google Scholar 

  • Clements JM, Roberts CF (1986) Transcription and processing signals in the 3-phosphoglycerate kinase (PGK) gene from Aspergillus nidulans. Gene 44:97–105

    Google Scholar 

  • Clutterbuck AJ (1974) Aspergillus nidulans genetics. In: King RC (ed) Handbook of genetics, vol 1. Plenum, New York, pp 447–510

    Google Scholar 

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

    Google Scholar 

  • Corrick CM, Twomey AP, Hynes MJ (1987) The nucleotide sequence of the amdS gene of Aspergillus nidulans and the molecular characterisation of 5′ mutations. Gene 53:63–71

    Google Scholar 

  • Cove DJ (1966) The induction and repression of nitrate reductase in the fungus Aspergillus nidulans. Biochim Biophys Acta 113:51–56

    Google Scholar 

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

    Google Scholar 

  • Der Garabedian PA, Lotti A-M, Vermeersch JJ (1986) 4-Aminobutyrate:2-oxoglutarate aminotransferase from Candida: purification and properties. Eur J Biochem 156:589–596

    Google Scholar 

  • Durrens P, Green PM, Arst HN, Scazzocchio C (1986) Heterologous insertion of transforming DNA and generation of new deletions associated with transformation in Aspergillus nidulans. Mol Gen Genet 203:544–549

    Google Scholar 

  • Forsburg SL, Guarente L (1988) Mutational analysis of upstream activation sequence 2 of the CYC1 gene of Saccharomyces cerevisiae: a HAP2-HAP3 responsive site. Mol Cell Biol 8:647–654

    Google Scholar 

  • Graves BJ, Johnson PF, McKnight SL (1986) Homologous recognition of a promoter domain common to the MSV LTR and the HSV tk gene. Cell 44:565–576

    Google Scholar 

  • Gwynne DI, Buxton FP, Sibley S, Davies RW, Lockington RA, Scazzocchio C, Sealy-Lewis HM (1987) Comparison of the cis-acting control regions of two coordinately controlled genes involved in ethanol utilisation in Aspergillus nidulans. Gene 51:205–216

    Google Scholar 

  • Hynes MJ (1975) Studies on the role of the areA gene in the regulation of nitrogen catabolism in Aspergillus nidulans. Aust J Biol Sci 28:301–313

    Google Scholar 

  • Hynes MJ (1978) Multiple independent control mechanisms affecting the acetamidase of Aspergillus nidulans. Mol Gen Genet 166:31–36

    Google Scholar 

  • Hynes MJ (1980) A mutation adjacent to gene amdS, defining the site of action of the positive control gene amdR in Aspergillus nidulans. J Bacteriol 142:400–406

    Google Scholar 

  • Hynes MJ, Corrick CM, King JA (1983) Isolation of genomic clones containing the amdS gene of Aspergillus nidulans and their use in the analysis of structural and regulatory mutations. Mol Cell Biol 3:1430–1439

    Google Scholar 

  • Hynes MJ, Corrick CM, Kelly JM, Littlejohn TG (1988) Identification of the sites of action of regulatory genes controlling the amdS gene of Aspergillus nidulans. Mol Cell Biol 8:2589–2596

    Google Scholar 

  • Kelly JM, Hynes MJ (1987) Multiple copies of the amdS gene of Aspergillus nidulans cause titration of trans-acting regulatory proteins. Curr Genet 12:21–31

    Google Scholar 

  • Kozak M (1986) Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell 44:283–292

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory. Cold Spring Harbor, New York

    Google Scholar 

  • Messing J (1983) New M13 vectors for cloning. Methods Enzymol 101c:20–78

    Google Scholar 

  • Norrander J, Kempe T, Messing J (1983) Construction of improved M13 vectors using oligodeoxynucleotide directed mutagenesis. Gene 26:101–106

    Google Scholar 

  • Olesen J, Hahn S, Guarente L (1987) Yeast HAP2 and HAP3 activators both bind to the CYC1 upstream activation site, UAS2, in an interdependent manner. Cell 51:953–961

    Google Scholar 

  • Pfeifer K, Prezant T, Guarente L (1987) Yeast HAP1 activator binds to two upstream activation sites of different sequences. Cell 49:19–27

    Google Scholar 

  • Pickett M, Gwynne DI, Buxton FP, Elliott R, Davies RW, Lockington RA, Scazzocchio C, Sealy-Lewis HM (1987) Cloning and characterisation of the aldA gene of Aspergillus nidulans. Gene 51:217–226

    Google Scholar 

  • Proudfoot N (1984) The end of the messenger and beyond. Nature 307:412–413

    Google Scholar 

  • Rambosek JA, Leach J (1986) Recombinant DNA in filamentous fungi: progress and prospects. CRC Crit Rev Biotechnol 6:357–391

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    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

    Google Scholar 

  • Upshall A (1986) Genetic and molecular characterisation of argB+ transformants of Aspergillus nidulans. Curr Genet 10:593–599

    Google Scholar 

  • Winkler H, Adam G, Mattes E, Schanz M, Hartig A, Ruis H (1988) Coordinate control of synthesis of mitochondrial and non-mitochondrial hemoproteins: a binding site for the HAP1 (CYC1) proteins in the UAS region of the yeast catalase T gene CTT1. EMBO J 7:1799–1804

    Google Scholar 

  • Yelton MM, Timberlake WE, van den Hondel CAMJJ (1985) A cosmid for selecting genes by complementation in Aspergillus nidulans: Selection of the developmentally regulated yA locus. Proc Natl Acad Sci USA 82:834–838

    Google Scholar 

  • Yonaha K, Suzuki K, Toyama S (1985) 4-Aminobutyrate:2-oxoglutarate aminotransferase of Streptomyces griseus: Purification and properties. Eur J Biochem 146:101–106

    Google Scholar 

Download references

Author information

Authors and Affiliations

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

    Imogen B. Richardson, Susan K. Hurley & Michael J. Hynes

Authors
  1. Imogen B. Richardson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Susan K. Hurley
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Michael J. Hynes
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by W. Gajewski

Rights and permissions

Reprints and permissions

About this article

Cite this article

Richardson, I.B., Hurley, S.K. & Hynes, M.J. Cloning and molecular characterisation of the amdR controlled gatA gene of Aspergillus nidulans . Mol Gen Genet 217, 118–125 (1989). https://doi.org/10.1007/BF00330950

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00330950

Key words

Search

Navigation