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ThehapC gene ofAspergillus nidulans is involved in the expression of CCAAT-containing promoters

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Abstract

The 5′ regulatory region of theamdS gene ofAspergillus nidulans, which encodes an acetamidase required for growth on acetamide as a carbon and nitrogen source, contains a CCAAT sequence which is required for setting the basal level ofamdS expression. Mobility shift studies have identified a factor inA. nidulans nuclear extracts which binds to this CCAAT sequence. InSaccharomyces cerevisiae theHAP3 gene encodes one component of a multisubunit complex that binds CCAAT sequences. A search of the EMBL and SwissProt databases has revealed anA. nidulans sequence with significant homology to theHAP3 gene adjacent to the previously cloned regulatory geneamdR. Sequencing of the remainder of this region has confirmed the presence of a gene, designatedhapC, with extensive homology toHAP3. The predicted amino acid sequence of HapC shows extensive identity to HAP3 in the central conserved domain, but shows little conservation in the flanking sequences. A haploid carrying ahapC deletion has been created and is viable, but grows poorly on all media tested. This null mutant grows especially slowly on acetamide as a sole carbon and nitrogen source, indicating thathapC plays a role inamdS expression. In agreement with this notion, it has been shown that thehapC deletion results in reduced levels of expression of anamdS::lacZ reporter gene and this effect is particularly evident under conditions of carbon limitation. Nuclear extracts prepared from thehapC deletion mutant show no CCAAT binding activity to theamdS orgatA promoters, indicating thathapC may encode a component of the complex binding at this sequence.

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References

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

    Google Scholar 

  • Andrianopoulos A, Hynes MJ (1990) Sequence and functional analysis of the positively acting regulatory geneamdR fromAspergillus nidulans. Mol Cell Biol 10:3194–3203

    Google Scholar 

  • Andrianopoulos A, Timberlake WE (1994) TheAspergillus nidulans abaA gene encodes a transcriptional activator that acts as a genetic switch to control development. Mol Cell Biol 14:2503–2515

    Google Scholar 

  • Aramayo R, Timberlake WE (1993) TheAspergillus nidulans yA gene is regulated byabaA. EMBO J 12:1039–2048

    Google Scholar 

  • Austin B, Hall RM, Tyler BM (1990) Optimization vectors and selection for transformation ofNeurospora crassa andAspergillus nidulans to bleomycin and phleomycin resistance. Gene 93:157–162

    Google Scholar 

  • Becker DM, Fikes JD, Guarente L (1991) A cDNA encoding a human CCAAT-binding protein cloned by functional complementation in yeast. Proc Natl Acad Sci USA 88:1968–1972

    Google Scholar 

  • Brent R, Ptashne M (1985) A eukaryotic transcriptional activator bearing the DNA specificity of a prokaryotic repressor. Cell 43:729–736

    Google Scholar 

  • Bucher P (1990) Weight matrix descriptions of four eukaryotic RNA polymerase II promoter elements derived from 502 unrelated promoter sequences. J Mol Biol 212:563–578

    Google Scholar 

  • Champe SP, Nagle DL, Yager LN (1994) Sexual sporulation. In: Martinelli SD, Kinghorn JR (eds)Aspergillus: 50 years on. Prog Indust Microbiol 29:429–454

    Google Scholar 

  • Clutterbuck AJ (1974)Aspergillus nidulans genetics. In: King RC (ed) Handbook of genetics, vol I. Plenum Press, New York, pp 265–273

    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 Press, Cold Spring Harbor, New York

    Google Scholar 

  • Corrick CM, Twomey AP, Hynes MJ (1987) The nucleotide sequence of theamdS gene ofAspergillus 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 fungusAspergillus nidulans. Biochem Biophys Acta 113:51–56

    Google Scholar 

  • Davidse LC (1986) Benzimidazole fungicides: mechanism of action and biological impact. Annu Rev Phytopathol 24:43–65

    Google Scholar 

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

    Google Scholar 

  • Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucl Acids Res 12:387–395

    Google Scholar 

  • Forsburg SL, Guarente L (1989) Identification and characterisation of HAP4: A third component of the CCAAT-bound HAP2/HAP3 heteromer. Genes Dev 3:1166–1178

    Google Scholar 

  • Hahn S, Guarente L (1988) Yeast HAP2 and HAP3: transcriptional activators in a heteromeric complex. Science 240:317–321

    Google Scholar 

  • Hahn S, Pinkham J, Wei R, Miller R, Guarente L (1988) TheHAP3 regulatory locus ofSaccharomyces cerevisiae encodes divergent overlapping transcripts. Mol Cell Biol 8:655–663

    Google Scholar 

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

    Google Scholar 

  • Hynes MJ (1980) A mutation, adjacent to geneamdS, defining the site of action of positive control geneamdR inAspergillus nidulans. J Bacteriol 142:400–406

    Google Scholar 

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

    Google Scholar 

  • Johnson PF, McKnight SL (1989) Eukaryotic transcriptional regulatory proteins. Ann Rev Biochem 58:799–839

    Google Scholar 

  • Li X, Mantovani R, Hooft van Huijsduijnen R, Andre I, Benoist C, Mathis D (1992) Evolutionary variation of the CCAAT-binding transcription factor NF-Y. Nucleic Acids Res 20:1087–1091

    Google Scholar 

  • Li Z, Kalassapudi SR, Childs G (1993) Isolation and characterisation of cDNAs encoding the sea urchin (Strongylocentropus purpuratus) homologue of the CCAAT binding protein NF-Y A subunit. Nucleic Acids Res 21:4639

    Google Scholar 

  • Littlejohn TG, Hynes MJ (1992) Analysis of the site of action of theamdR product for the regulation of theamdS gene ofAspergillus nidulans. Mol Gen Genet 235:81–88

    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 

  • McNabb DS, Xing Y, Guarente L (1995) Cloning of yeastHAP5: a novel subunit of a heteromeric complex required for CCAAT binding. Genes Dev 9:47–58

    Google Scholar 

  • Mulder W, Scholten IHJM, de Boer RW, Grivell LA (1994) Sequence of the HAP3 transcription factor ofKluyveromyces lactis predicts the presence of a novel 4-cysteine zinc-finger motif. Mol Gen Genet 245:96–106

    Google Scholar 

  • Nagata O, Takashima T, Tanaka M, Tsukagoshi N (1993)Aspergillus nidulans nuclear proteins bind to a CCAAT element and the adjacent upstream sequence in the promoter region of the starch-inducible Taka-amylase A gene. Mol Gen Genet 237:251–260

    Google Scholar 

  • Nguyen C, Bolotin-Fukuhara M, Wésolowski-Louvel M, Fukuhara H (1995) The respiratory system ofKluyveromyces lactis escapes from HAP2 control. Gene 152:113–115

    Google Scholar 

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

    Google Scholar 

  • Olesen JT, Guarente L (1990) The HAP2 subunit of yeast CCAAT transcriptional activator contains adjacent domains for subunit association and DNA recognition: model for the HAP2/3/4 complex. Genes Dev 4:1714–1729

    Google Scholar 

  • Olesen JT, Fikes JD, Guarente L (1991) TheSchizosaccharomyces pombe homolog ofSaccharomyces cerevisiae HAP2 reveals selective and stringent conservation of the small essential core protein domain. Mol Cell Biol 11:611–619

    Google Scholar 

  • Pinkham J, Guarente L (1985) Cloning and molecular analysis of theHAP2 locus: a global regulator of respiratory genes inSaccharomyces cerevisiae. Mol Cell Biol 5:3410–3416

    Google Scholar 

  • Ptashne M (1988) How eukaryotic transcriptional activators work. Nature 335:683–689

    Google Scholar 

  • Richardson IB, Katz ME, Hynes MJ (1992) Molecular characterisation of thelam locus and sequences involved in regulation by the AmdR protein ofAspergillus nidulans. Mol Cell Biol 12:337–346

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  • Sinha S, Maity SN, Lu J, de Crombrugghe B (1995) Recombinant rat CBF-C, the third subunit of CBF/NFY, allows formation of a protein-DNA complex with CBF-A and CBF-B and with yeast HAP2 and HAP3. Proc Natl Acad Sci USA 92:1624–1628

    Google Scholar 

  • Van Heeswijck R, Hynes MJ (1991) TheamdR product and a CCAAT-binding factor bind to adjacent, possibly overlapping DNA sequences in the promoter region of theAspergillus nidulans amdS gene. Nucleic Acids Res 19:2655–2660

    Google Scholar 

  • Xing Y, Fikes JD, Guarente L (1993) Mutations in yeast HAP2/HAP3 define a hybrid CCAAT box binding domain. EMBO J 12:4647–4655

    Google Scholar 

  • Zitomer RS, Lowry RS (1992) Regulation of gene expression by oxygen inSaccharomyces cerevisiae. Microbiol Rev 50:1–11

    Google Scholar 

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Communicated by E. Cerdá-Olmedo

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Papagiannopoulos, P., Andrianopoulos, A., Sharp, J.A. et al. ThehapC gene ofAspergillus nidulans is involved in the expression of CCAAT-containing promoters. Molec. Gen. Genet. 251, 412–421 (1996). https://doi.org/10.1007/BF02172369

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  • DOI: https://doi.org/10.1007/BF02172369

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