Summary
We have determined the structure and complete nucleotide sequence of the trifunctional trpC gene from the Ascomycetous fungus Aspergillus nidulans. Results from RNA gel blot analyses showed that this gene encodes two size classes of polyribosomal, poly (A)+RNAs with approximate lengths of 2,400 and 2,600 nucleotides. S1 nuclease protection studies demonstrated that the distribution into the two size classes is due to selection of alternative sites for polyadenylation. The transcription units contain a single open translation reading frame of 2,304 nucleotides. The sequence of this reading frame is approximately 40% divergent from the sequence of the functionally analogous trp-1 gene from Neurospora crassa (Schechtman, M.G. and Yanofsky, C., J. Mol. Appl. Gen. 2:83–99). The predicted amino acid sequence of the A. nidulans trpC polypeptide is also 40% divergent from the predicted amino acid sequence of the N. crassa trp-1 polypeptide. The A. nidulans gene has considerably less bias in codon selection than observed for the N. crassa gene. Discrete regions of DNA homology were also found in similar positions in the 5′ and 3′ flanking sequences of the Aspergillus and Neurospora genes. Similar regions of homology were not observed in other Aspergillus or Neurospora genes that have been sequenced. Thus, if these evolutionarily conserved sequences act as signals for transcription initiation or polyadenylation, or are involved in gene regulation, their functions are restricted to a subset of protein coding genes in these two closely related fungi.
Similar content being viewed by others
References
Alton NK, Buxton F, Patel V, Giles NH, Vapnek D (1982) 5′ untranslated sequences of two structural genes in the qa gene cluster of Neurospora crassa. Proc. Natl. Acad. Sci. USA 79:1955–1959
Arends H, Sebald W (1984) Nucleotide sequence of the cloned mRNA and gene of the ADP/ATP carrier from Neurospora crassa. EMBO J 3:377–382
Arst HN (Jr) (1981) Aspects of the control of gene expression in fungl. Sym. Soc. Gen. Microbiol. 31:131–160
Ballance DJ, Buxton FP, Turner G (1983) Transformation of Aspergillus nidulans by the orotidine-5′-phosphate decarboxylase gene of Neurospora crassa. Biochem. Biophys. Res. Commun. 112:284–289
Berget SM (1984) Are U4 small ribonucleoproteins involved in polyadenylation? Nature 309:179–182
Bergen LG, Upshall A, Morris NR (1984) S-phase, G2, and nuclear division mutants of Aspergillus nidulans. J. Bacteriol. 159:114–119
Berk AJ, Sharp PA (1977) Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1-endonuclease digested hybrids. Cell 12:721–732
Berse B, Dmochowska A, Skrzypek M, Weglenski P, Bates MA, Weiss RL (1983) Cloning and characterization of the ornithine carbamoyl-transferase gene from Aspergillus nidulans. Gene 25:109–117
Boel E, Hansen MT, Hjort I, Hoegh I, Fiil NP (1984) Two different types of intervening sequences in the glucoamylase gene from Aspergillus niger. 3:1581–1585
Boyer HW, Roulland-Dussoix D (1969) A complementation analysis of the restriction and modification of DNA in Escherichia coli. J. Mol. Biol 41:459–472
Breathnach R, Chambon P (1981) Organization and expression of eukaryotic split genes coding for proteins. Ann Rev. Biochem 50:349–383
Burke RL, Tekamp-Olsen P, Najarian R (1983) The isolation, characterization, and sequence of the pyruvate kinase gene of Sacharomyces cerevisiae. J. Biol. Chem. 258:2193–2201
Champe S, Kurtz MB, Yager LN, Butrick NJ, Axelrod DE (1981) Spore formation in Aspergillus nidulans: Competence and other developmental processes. In Turian G, Hohl H (eds) The fungal spore: morphogenetic controls. Academic Press, New York, p 255
Clewell DB, Helinski DR (1969) Effect of growth conditions on the formation of the relaxation complex of supercoiled ColE1 deoxyribonucleic acid and protein in Escherichia coli. J. Bacteriol 110:1135–1148
Clutterbuck AJ (1974) Aspergillus nidulans. In King RC (ed) Handbook of genetics, vol I. Plenum Press, New York, p 447
Clutterbuck AJ (1977) The genetics of conidiation of Aspergillus nidulans. In Smith JE, Pateman JA (eds) Genetics and physiology of Aspergillus. Academic Press, New York, p 305
Cove DJ (1979) Genetic studies of nitrate assimilation in Aspergillus nidulans. Biol. Rev. 54: 291–327
Goldberg M (1979) Ph. D. Thesis. Stanford University, Stanford, California
Gwynne DI, Miller BL, Miller KY, Timberlake WE (1984) Structure and regulated expression of the SpoCl gene cluster from Aspergillus nidulans. J. Mol. Biol. 179 (in press)
Hulett FM, DeMoss JA (1975) Subunit structure of anthranilate synthetase from Neurospora crassa. J. Biol. Chem. 250: 6648–6652
Hutter R, DeMoss JA (1967) Enzyme analysis of the tryptophan pathway in Aspergillus nidulans. Genetics 55:241–247
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
Jackson AO, Larkins BA (1976) Influence of ionic strength, pH, and chelation of divalent metals on isolation of polyribosomes from tobacco leaves. Plant Physiol. 57:5–10
Käfer E (1977) The anthranilate synthetase enzyme complex and the trifunctional trpC gene of Aspergillus. Can. J. Genet. Cytol. 19:723–738
Kinnaird JH, Fincham JRS (1983) The complete nucleotide sequence of the Neurospora crassa am (NADP-specific glutamate dehydrogenase) gene. Gene 26:253–260
Lehrach H, Diamond D, Wozney JM, Boedtker H (1977) RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry 16:4743–4750
Maniatis T, Jeffrey A, Kleid D (1975) Nucleotide sequence of the rightward operator of phage λ. Proc. Natl. Acad. Sci. USA 72:1184–1188
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbour Laboratory. Cold Spring Harbour, New York, p 86
Maxam AM, Gilbert W (1980) Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol 65:499–560
Messing J (1983) New M13 vectors for cloning. Methods. Enzymol. 100B:20–78
Morris N (1976) Mitotic mutants of Aspergillus nidulans. Genet. Res. 26:237–254
Morris N, Lai M, Oakley CE (1979) Identification of a gene for F-tubulin in Aspergillus nidulans. Cell 16:437–442
Morris N, Kirsch D, Oakley BR (1982) Molecular and genetic methods for studying mitosis and spindle proteins in Aspergillus nidulans. Methods Cell Biol. 25:107–130
Oakley BR, Morris NR (1980) Nuclear movement is F-tubulin dependent in Aspergillus nidulans. Cell 19:255–262
Orr W, Timberlake WE (1982) Clustering of spore-specific genes in Aspergillus nidulans. Proc. Natl. Acad. Sci. USA 79: 5976–5980
Poncz M, Solowiejczyk D, Ballantine M, Schwartz E, Surrey S (1982) “Nonrandom” DNA sequence analysis in bacteriophage M13 by the dideoxy chain-termination method. Proc. Natl. Acad. Sci. USA 79:4298–4302
Pontecorvo G, Roper JA, Hemmons ZM, MacDonald KD, Bufton AWJ (1953) The genetics of Aspergillus nidulans. Adv. Gen. 5:141–238
Proudfoot NJ, Brownlee GG (1976) 3′ non-coding region sequence in eukaryotic messenger RNA. Nature 263:211–214
Rigby PWJ, Dieckman M, Rhodes C, Berg P (1977) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J. Molec. Biol 113: 237–251
Roberts CF (1967) Complementation analysis of the tryptophan pathway in Aspergillus nidulans. Genetics 55:233–239
Rozek CE, Orr WE, Timberlake WE (1978) Diversity and abundance of polyadenylated RNA from Achlya ambisexualis. Biochemistry 17:716–722
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-termination inhibitors. Proc. Natl. Acad. Sci. USA 74: 5463–5467
Schechtman MG, Yanofsky C (1983) Structure of the trifunctional trp-1 gene of Neurospora crassa and its aberrent expression in Escherichia coli. J. Molec. Appl. Genet 2:83–99
Tilburn J, Scazzocchio C, Taylor GG, Zabicky-Zissman JH, Lockington RA, Davies RW (1984) Transformation by integration in Aspergillus nidulans. Gene 26: 205–221
Timberlake WE (1976) Alterations in RNA and protein synthesis associated with steroid hormone-induced sexual morphogenesis in the water mold Achlya. Dev. Biol. 51:202–214
Timberlake WE (1980) Developmental gene regulation in Aspergillus nidulans. Dev. Biol. 78:497–510
Timberlake WE, Barnard EL (1981) Organization of a gene cluster expressed specifically in the asexual spores of A. nidulans. Cell 26:29–37
Timberlake WE, Gwynne DI, Hamer JE, Miller BL, Miller KY, Mullaney EM, deSouza ER, Yelton MM, Zimmerman CR (1983) Gene regulation during conidiation in Aspergillus nidulans. In Goldberg RB (ed) Plant molecular biology. Alan R. Liss, New York, p 179
Yelton MM, Hamer JE, deSouza ER, Mullaney EJ, Timberlake WE (1983) Developmental regulation of the Aspergillus nidulans trpC gene. Proc. Natl. Acad. Sci. USA 80:7576–7580
Yelton MM, Hamer JE, Timberlake WE (1984a) Transformation of Aspergillus nidulans by using a trpC plasmid. Proc. Natl. Acad. Sci. USA 81:1470–1474
Yelton MM, Timberlake WE, van den Hondel CAMJJ (1984b) A cosmid for selecting genes by complementation in Aspergillus: Selection of the developmentally regulated yA locus. Proc. Natl. Acad. Sci. USA (in press)
Author information
Authors and Affiliations
Additional information
Communicated by R.B. Goldberg
Rights and permissions
About this article
Cite this article
Mullaney, E.J., Hamer, J.E., Roberti, K.A. et al. Primary structure of the trpC gene from Aspergillus nidulans . Molec Gen Genet 199, 37–45 (1985). https://doi.org/10.1007/BF00327506
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00327506