Summary
Three alcohol dehydrogenase (ADH) genes have recently been characterized in the yeast Kluyveromyces lactis. We report on a fourth ADH in K. lactis (KADH II: KADH2 Footnote 1 gene) which is highly similar to other ADHs in K. lactis and Saccharomyces cerevisiae. KADH II appears to be a cytoplasmic enzyme, and after expression of KADH2 in S. cerevisiae enzyme activity comigrated with a K. lactis ADH present in cells grown in glucose or in ethanol. KADH I was also expressed in S. cerevisiae and it comigrated with a major ADH species expressed under glucose growth conditions in K. lactis. The substrate specificities for KADH I and KADH II were shown to be more similar to that of SADH II than to SADH I. SADH I cannot efficiently utilize long chain alcohols, in contrast to other cytoplasmic yeast ADHs, presumably because of the presence of a methionine (residue 271) in its substrate binding cleft. A comparison of the DNA sequences of ADHs among K. lactis, S. cerevisiae and Schizosaccharomyces pombe suggests that the ancestral yeast species contained one cytoplasmic ADH. After divergence from S. pombe, the ADH in the ancestor to K. lactis and S. cerevisiae was duplicated, and one ADH became localized to the mitochondrion, presumably for the oxidative use of ethanol. Following the speciation of S. cerevisiae and K. lactis, the gene encoding the cytoplasmic ADH in S. cerevisiae duplicated, which resulted in the development of the SADH II protein as the primary oxidative enzyme in place of SADH III. In contrast, the K. lactis mitochondrial ADH duplicated to give rise to the highly expressed KADH3 and KADH4 genes, both of which may still play primary roles in oxidative metabolism. These data suggest that K. lactis and S. cerevisiae use different compartments for their metabolism of ethanol. Our results also indicate that the complex regulatory circuits controlling the glucose-repressible SADH2 in S. cerevisiae are a recent acquisition from regulatory networks used for the control of genes other than SADH2.
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Notes
The EMBL Library Accession Number for KADH2 is: X64397
References
Bennetzen JL, Hall BD (1982) The primary structure of the Saccharomyces cerevisiae gene for alcohol dehydrogenase 1. J Biol Chem 257:3018–3025
Ciriacy M (1975) Genetics of alcohol dehydrogenase II in Saccharomyces cerevisiae. Two loci controlling synthesis of the glucose repressible ADH II. Mol Gen Genet 138:157–164
Denis CL, Young ET (1983) Isolation and characterization of the positive regulatory gene ADR1 from Saccharomyces cerevisiae. Mol Cell Biol 3:360–370
Denis CL, Ferguson J, Young ET (1983) mRNA levels for the fermentative alcohol dehydrogenase of Saccharomyces cerevisiae decrease upon growth on a nonfermentable carbon source. J Biol Chem 258:1165–1171
Doolittle RE (1981) Similar amino acid sequences: Chance or common ancestry? Science 214:149–158
Drewke C, Ciriacy M (1988) Overexpression, purification and properties of alcohol dehydrogenase IV from Saccharomyces cerevisiae. Biochim Biophys Acta 950:54–60
Eklund H, Muller-Wille P, Horjales E, Futer O, Holmquist B, Vallee BL, Hoog J, Kaiser R, Jornvall H (1990) Comparison of three classes of human liver alcohol dehydrogenase. Eur J Biochem 193:303–310
Gregoret LM, Rader SD, Fletterick RJ, Cohen FE (1991) Hydrogen bonds involving sulfur atoms in proteins. Proteins 9:99–107
Grieve PA, Kitchen BJ, Dulley JR (1983) Partial characterization of cheese-ripening proteinases produced by the yeast Kluyveromyces lactis. J Dairy Res 50:469–480
Irwin DM, Wilson AC (1990) Concerted evolution of ruminant stomach lysozymes. J Biol Chem 265:4944–4952
Ito H, Fukada Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168
Jornvall H, Persson B, Jeffrey J (1987) Characteristics of alcohol/polyol dehydrogenases. Eur J Biochem 167:195–201
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, New York
Paquin CE, Williamson VM (1986) Ty insertions at two loci account for most of the spontaneous antimycin A resistance mutations during growth at 15° C of Saccharomyces cerevisiae strains lacking ADH1. Mol Cell Biol 6:70–79
Russell DW, Smith M, Williamson VM, Young ET (1983) Nucleotide sequence of the yeast alcohol dehydrogenase II gene. J Biol Chem 258:2674–2682
Russell PR, Hall BD (1983) The primary structure of the alcohol dehydrogenase gene from the fission yeast Schizosaccharomyces pombe. J Biol Chem 258:143–149
Saliola M, Shuster JR, Falcone C (1990) The alcohol dehydrogenase system in the yeast Kluyveromyces lactis. Yeast 6:193–204
Saliola M, Gonnella R, Mazzoni C, Falcone C (1991) Isolation of two genes encoding putative mitochondrial alcohol dehydrogenases in the yeast Kluyveromyces lactis. Yeast 1:391–401
Salmeron JM, Johnston SA (1986) Analysis of the Kluyveromyces lactis positive regulatory gene LAC9 reveals functional homology to, but sequence divergence from, the Saccharomyces cerevisiae GAL4 gene. Nucleic Acids Res 14:7767–7781
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Scherer S, Davis RW (1980) Recombination of dispersed repeated DNA sequences in yeast. Science 209:1380–1384
Simon M, Adam G, Raputz W, Spevak W, Ruis H (1991) The Saccharomyces cerevisiae ADR1 gene is a positive regulator of transcription of genes encoding peroxisomal proteins. Mol Cell Biol 11:699–704
Swofford DL (1989) PAUP: phylogenetic analysis using parsimony, version 3.0b. Illinois Natural History Survey, Champlain
Verduyn C, Breedveld GJ, Scheffers WA, Van Dijken JP (1988) Substrate specificity of alcohol dehydrogenase from the yeasts Hansenula polymorpha CBS 4732 and Candida utilis CBS 621. Yeast 4:143–148
Weisenfeld M, Schimpfessel I, Crokaert R (1975) Multiple forms of mitochondrial alcohol dehydrogenase in Saccharomyces cerevisiae. Biochim Biophys Acta 405:500–512
Williamson VM, Paquin CE (1987) Homology of Saccharomyces cerevisiae ADH4 to an iron-activated alcohol dehydrogenase from Zymomonas mobilis. Mol Gen Genet 209:374–381
Williamson VM, Bennetzen J, Young ET, Nasmyth K, Hall BD (1980) Isolation of the structural gene for alcohol dehydrogenase by genetic complementation in yeast. Nature 283:214–216
Wills C (1976) Production of yeast alcohol dehydrogenase isozymes by selection. Nature 261:26–29
Wills C, Jornvall H (1979) The two major isozymes of yeast alcohol dehydrogenases. Eur J Biochem 99:323–331
Wray LV, Witte MM, Dickson RC, Riley MI (1987) Characterization of a positive regulatory gene, LAC9, that controls induction of the lactose-galactose regulon of Kluyveromyces lactis: Structural and functional relationships to GAL4 of Saccharomyces cerevisiae. Mol Cell Biol 7:1111–1121
Young ET, Pilgrim D (1985) Isolation and DNA sequence of ADH3, a nuclear gene encoding the mitochondrial isozyme of alcohol dehydrogenase in Saccharomyces cerevisiae. Mol Cell Biol 5:3024–3034
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Communicated by C.P. Hollenberg
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Shain, D.H., Salvadore, C. & Denis, C.L. Evolution of the alcohol dehydrogenase (ADH) genes in yeast: characterization of a fourth ADH in Kluyveromyces lactis . Molec. Gen. Genet. 232, 479–488 (1992). https://doi.org/10.1007/BF00266253
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DOI: https://doi.org/10.1007/BF00266253