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. 1985 Aug;4(8):2093–2099. doi: 10.1002/j.1460-2075.1985.tb03897.x

Identification and molecular analysis of a third Aspergillus nidulans alcohol dehydrogenase gene.

G L McKnight, H Kato, A Upshall, M D Parker, G Saari, P J O'Hara
PMCID: PMC554467  PMID: 2998782

Abstract

An Aspergillus nidulans functional cDNA encoding an alcohol dehydrogenase (ADH) was isolated by its ability to complement an adh1 mutation in Saccharomyces cerevisiae. Alignment of the cDNA and cloned genomic DNA sequences indicated that the ADH gene contains two small introns. The presence of ethanol in the growth medium was shown to result in ADH mRNA accumulation presumably due to transcriptional induction of the gene. However, ADH mRNA accumulation was at most only partially repressed by the presence of glucose. The ADH gene characterized here is designated ADH3 since it is distinct from the alcA gene which encodes ADH I and appears distinct from the gene which encodes ADH II. We demonstrated that the first intron in the A. nidulans ADH3 gene was not efficiently spliced in S. cerevisiae whereas the promoter region was utilized weakly. We also present a comparison of the primary structure of A. nidulans ADH III with the alcohol dehydrogenases of S. cerevisiae and Schizosaccharomyces pombe.

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Selected References

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  1. Ballance D. J., Buxton F. P., Turner G. Transformation of Aspergillus nidulans by the orotidine-5'-phosphate decarboxylase gene of Neurospora crassa. Biochem Biophys Res Commun. 1983 Apr 15;112(1):284–289. doi: 10.1016/0006-291x(83)91828-4. [DOI] [PubMed] [Google Scholar]
  2. Beggs J. D. Transformation of yeast by a replicating hybrid plasmid. Nature. 1978 Sep 14;275(5676):104–109. doi: 10.1038/275104a0. [DOI] [PubMed] [Google Scholar]
  3. Bennetzen J. L., Hall B. D. Codon selection in yeast. J Biol Chem. 1982 Mar 25;257(6):3026–3031. [PubMed] [Google Scholar]
  4. Bennetzen J. L., Hall B. D. The primary structure of the Saccharomyces cerevisiae gene for alcohol dehydrogenase. J Biol Chem. 1982 Mar 25;257(6):3018–3025. [PubMed] [Google Scholar]
  5. Berget S. M. Are U4 small nuclear ribonucleoproteins involved in polyadenylation? Nature. 1984 May 10;309(5964):179–182. doi: 10.1038/309179a0. [DOI] [PubMed] [Google Scholar]
  6. Berse B., Dmochowska A., Skrzypek M., Wegleński P., Bates M. A., Weiss R. L. Cloning and characterization of the ornithine carbamoyltransferase gene from Aspergillus nidulans. Gene. 1983 Nov;25(1):109–117. doi: 10.1016/0378-1119(83)90173-7. [DOI] [PubMed] [Google Scholar]
  7. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Boel E., Hansen M. T., Hjort I., Høegh I., Fiil N. P. Two different types of intervening sequences in the glucoamylase gene from Aspergillus niger. EMBO J. 1984 Jul;3(7):1581–1585. doi: 10.1002/j.1460-2075.1984.tb02014.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Brändén C. I., Eklund H., Cambillau C., Pryor A. J. Correlation of exons with structural domains in alcohol dehydrogenase. EMBO J. 1984 Jun;3(6):1307–1310. doi: 10.1002/j.1460-2075.1984.tb01967.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Casadaban M. J., Cohen S. N. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. doi: 10.1016/0022-2836(80)90283-1. [DOI] [PubMed] [Google Scholar]
  11. Ciriacy M. Isolation and characterization of further cis- and trans-acting regulatory elements involved in the synthesis of glucose-repressible alcohol dehydrogenase (ADHII) in Saccharomyces cerevisiae. Mol Gen Genet. 1979 Nov;176(3):427–431. doi: 10.1007/BF00333107. [DOI] [PubMed] [Google Scholar]
  12. Creaser E. H., Porter R. L., Britt K. A., Pateman J. A., Doy C. H. Purification and preliminary characterization of alcohol dehydrogenase from Aspergillus nidulans. Biochem J. 1985 Jan 15;225(2):449–454. doi: 10.1042/bj2250449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dagert M., Ehrlich S. D. Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene. 1979 May;6(1):23–28. doi: 10.1016/0378-1119(79)90082-9. [DOI] [PubMed] [Google Scholar]
  14. Doy C. H., Pateman J. A., Olsen J. E., Kane H. J., Creaser E. H. Genomic clones of Aspergillus nidulans containing alcA, the structural gene for alcohol dehydrogenase and alcR, a regulatory gene for ethanol metabolism. DNA. 1985 Apr;4(2):105–114. doi: 10.1089/dna.1985.4.105. [DOI] [PubMed] [Google Scholar]
  15. Eklund H., Brändén C. I., Jörnvall H. Structural comparisons of mammalian, yeast and bacillar alcohol dehydrogenases. J Mol Biol. 1976 Mar 25;102(1):61–73. doi: 10.1016/0022-2836(76)90073-5. [DOI] [PubMed] [Google Scholar]
  16. Henikoff S., Kelly J. D., Cohen E. H. Transcription terminates in yeast distal to a control sequence. Cell. 1983 Jun;33(2):607–614. doi: 10.1016/0092-8674(83)90441-5. [DOI] [PubMed] [Google Scholar]
  17. Innis M. A., Holland M. J., McCabe P. C., Cole G. E., Wittman V. P., Tal R., Watt K. W., Gelfand D. H., Holland J. P., Meade J. H. Expression, Glycosylation, and Secretion of an Aspergillus Glucoamylase by Saccharomyces cerevisiae. Science. 1985 Apr 5;228(4695):21–26. doi: 10.1126/science.228.4695.21. [DOI] [PubMed] [Google Scholar]
  18. Jörnvall H. Differences between alcohol dehydrogenases. Structural properties and evolutionary aspects. Eur J Biochem. 1977 Feb;72(3):443–452. doi: 10.1111/j.1432-1033.1977.tb11268.x. [DOI] [PubMed] [Google Scholar]
  19. Keller E. B., Noon W. A. Intron splicing: a conserved internal signal in introns of animal pre-mRNAs. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7417–7420. doi: 10.1073/pnas.81.23.7417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Langford C., Nellen W., Niessing J., Gallwitz D. Yeast is unable to excise foreign intervening sequences from hybrid gene transcripts. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1496–1500. doi: 10.1073/pnas.80.6.1496. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lockington R. A., Sealy-Lewis H. M., Scazzocchio C., Davies R. W. Cloning and characterization of the ethanol utilization regulon in Aspergillus nidulans. Gene. 1985;33(2):137–149. doi: 10.1016/0378-1119(85)90088-5. [DOI] [PubMed] [Google Scholar]
  22. McKnight G. L., McConaughy B. L. Selection of functional cDNAs by complementation in yeast. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4412–4416. doi: 10.1073/pnas.80.14.4412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mullaney E. J., Hamer J. E., Roberti K. A., Yelton M. M., Timberlake W. E. Primary structure of the trpC gene from Aspergillus nidulans. Mol Gen Genet. 1985;199(1):37–45. doi: 10.1007/BF00327506. [DOI] [PubMed] [Google Scholar]
  24. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  25. Nunberg J. H., Meade J. H., Cole G., Lawyer F. C., McCabe P., Schweickart V., Tal R., Wittman V. P., Flatgaard J. E., Innis M. A. Molecular cloning and characterization of the glucoamylase gene of Aspergillus awamori. Mol Cell Biol. 1984 Nov;4(11):2306–2315. doi: 10.1128/mcb.4.11.2306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pateman J. A., Doy C. H., Olsen J. E., Norris U., Creaser E. H., Hynes M. Regulation of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (AldDH) in Aspergillus nidulans. Proc R Soc Lond B Biol Sci. 1983 Feb 22;217(1208):243–264. doi: 10.1098/rspb.1983.0009. [DOI] [PubMed] [Google Scholar]
  27. Russell D. W., Smith M., Williamson V. M., Young E. T. Nucleotide sequence of the yeast alcohol dehydrogenase II gene. J Biol Chem. 1983 Feb 25;258(4):2674–2682. [PubMed] [Google Scholar]
  28. Russell P. R. Evolutionary divergence of the mRNA transcription initiation mechanism in yeast. Nature. 1983 Jan 13;301(5896):167–169. doi: 10.1038/301167a0. [DOI] [PubMed] [Google Scholar]
  29. Russell P. R., Hall B. D. The primary structure of the alcohol dehydrogenase gene from the fission yeast Schizosaccharomyces pombe. J Biol Chem. 1983 Jan 10;258(1):143–149. [PubMed] [Google Scholar]
  30. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  32. Teem J. L., Abovich N., Kaufer N. F., Schwindinger W. F., Warner J. R., Levy A., Woolford J., Leer R. J., van Raamsdonk-Duin M. M., Mager W. H. A comparison of yeast ribosomal protein gene DNA sequences. Nucleic Acids Res. 1984 Nov 26;12(22):8295–8312. doi: 10.1093/nar/12.22.8295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tilburn J., Scazzocchio C., Taylor G. G., Zabicky-Zissman J. H., Lockington R. A., Davies R. W. Transformation by integration in Aspergillus nidulans. Gene. 1983 Dec;26(2-3):205–221. doi: 10.1016/0378-1119(83)90191-9. [DOI] [PubMed] [Google Scholar]
  35. Timberlake W. E., Barnard E. C. Organization of a gene cluster expressed specifically in the asexual spores of A. nidulans. Cell. 1981 Oct;26(1 Pt 1):29–37. doi: 10.1016/0092-8674(81)90030-1. [DOI] [PubMed] [Google Scholar]
  36. Williamson V. M., Bennetzen J., Young E. T., Nasmyth K., Hall B. D. Isolation of the structural gene for alcohol dehydrogenase by genetic complementation in yeast. Nature. 1980 Jan 10;283(5743):214–216. doi: 10.1038/283214a0. [DOI] [PubMed] [Google Scholar]
  37. Wills C. Production of yeast alcohol dehydrogenase isoenzymes by selection. Nature. 1976 May 6;261(5555):26–29. doi: 10.1038/261026a0. [DOI] [PubMed] [Google Scholar]
  38. Yelton M. M., Hamer J. E., Timberlake W. E. Transformation of Aspergillus nidulans by using a trpC plasmid. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1470–1474. doi: 10.1073/pnas.81.5.1470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Yelton M. M., Hamer J. E., de Souza E. R., Mullaney E. J., Timberlake W. E. Developmental regulation of the Aspergillus nidulans trpC gene. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7576–7580. doi: 10.1073/pnas.80.24.7576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Yelton M. M., Timberlake W. E., Hondel C. A. A cosmid for selecting genes by complementation in Aspergillus nidulans: Selection of the developmentally regulated yA locus. Proc Natl Acad Sci U S A. 1985 Feb;82(3):834–838. doi: 10.1073/pnas.82.3.834. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Zaret K. S., Sherman F. DNA sequence required for efficient transcription termination in yeast. Cell. 1982 Mar;28(3):563–573. doi: 10.1016/0092-8674(82)90211-2. [DOI] [PubMed] [Google Scholar]

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