Gene Dmel\Adh

Structural gene for alcohol dehydrogenase [ADH (EC 1.1.1.1)]. Natural populations are polymorphic for three electrophoretic alleles (AdhF, AdhS, AdhF-ChD) and for three rarer electrophoretic alleles (AdhUS, AdhF', AdhUF). The frequency of the AdhF allele increases, at the expense of AdhS, with increasing latitude in both northern and southern hemispheres [Johnson and Schaffer, 1973, Biochem. Genet. 10: 149-63; Vigue and Johnson, 1973, Biochem. Genet. 9: 213-27; Wilks, Gibson, Oakeshott and Chambers, 1980, Aust. J. Biol. Sci. 33: 575-85; Anderson, 1981, Genetic Studies of Drosophila Populations (Gibson and Oakes, eds.). Australian National University Press, pp. 237-50; Anderson and Chambers, 1982, Evolution 36: 86-96]. Confers resistance to ethanol; flies lacking ADH rapidly become intoxicated and eventually die on exposure to ethanol (Grell, Jacobson and Murphy, 1968, Ann. N.Y. Acad. Sci 151: 441-45; Vigue and Sofer, 1976, Biochem. Genet. 14: 127-135; David, Bocquet, Arens and Fouillet, 1976, Biochem. Genet. 14: 989-97). However, ethanol sensitivity is complex since even Adh nulls are more resistant to ethanol when young than when old (Vigue and Sofer, 1976; Tsubota). Adh+ flies are killed by low concentrations of unsaturated secondary alcohols (e.g. 1-penten-3-ol; 1-pentyn-3-ol) but not by unsaturated primary alcohols (e.g. 1-penten-1-ol) (Sofer and Hatkoff, 1972, Genetics 72: 545-49), presumably due to the formation of toxic ketones. This allows the chemical selection of Adh nulls (Sofer and Hatkoff, 1972; O'Donnell, Gerace, Leister and Sofer, 1975, Genetics 79: 73-83). ADH may play a metabolic role independent of alcohol detoxication, i.e. in the metabolism of higher alcohols (see Winberg, Thatcher and McKinley-McKee, 1982, Biochem. Biophys. Acta 704: 7-16). ADH also catalyses the oxidation of acetaldehyde to acetate (Heinstra, Eisses, Schoonen, Aben, de Winter, van de Horst, van Marrewijk, Beenakkers, Scharloo and Thorig, 1983, Genetica 60: 129-37; Moxon, Holmes, Parsons, Irving, and Doddrell, 1985, Comp. Biochem. Physiol. 80B: 525-35). Specific activity of ADH changes with development, with peaks at the end of the third larval instar and about four days after eclosion (Ursprung, Sofer and Burroughs, 1970, Wilhelm Roux's Arch. Entwicklungsmech. Org. 164: 201-08; Dunn, Wilson and Jacobson, 1969, J. Exp. Zool. 171: 185-90; Leibenguth, Rammo and Dubiczky, 1979, Wilhelm Roux's Arch. Dev. Biol. 187: 81-88; Maroni and Stamey, 1983, DIS 59: 77-79; Anderson and McDonald, 1981, Canad. J. Genet. Cytol. 23: 305-13). Most of the activity is in the larval fat body and gut and the adult fat body (Ursprung, Sofer and Burroughs). Maternal inheritance of ADH by embryos and larvae (O'Donnell et al.; Leibenguth et al.). Half life of ADH-F in vivo estimated as 55.3 hours (Anderson and McDonald, 1981, Biochem. Genet. 19: 411-19). Not expressed in SL2 tissue culture cells, but transfected cloned gene is (Benyajati and Dray, 1984, Proc. Nat. Acad. Sci. 1701-05). Ethanol tolerance usually correlated with ADH activity and polymorphic experimental populations exposed to ethanol usually show an increase in the frequency AdhF (McDonald and Avise, 1976, Biochem. Genet. 14: 347-55; Cavener and Clegg, 1978, Genetics 90: 629-44; van Delden, Kamping and van Dijk, 1975, Experientia 31: 418-19; Oakeshott, Gibson, Anderson and Champ, 1980, Aust. J. Biol. Sci. 33: 105-14; McDonald, Chambers, David and Ayala, 1977, Proc. Nat. Acad. Sci. USA 74: 4562-66). Flies carrying AdhF tend to be more resistant than those carrying only AdhS to ethanol [Kamping and van Delden, 1978, Biochem. Genet. 16: 541-55; Ainsley and Kitto, 1975, Isozymes (C. Markert, ed.). Academic Press, Vol. II, pp. 733-43; Briscoe, Robertson and Malpica, 1975, Nature (London) 253: 148-49]. Electrophoresis of homozygous genotypes usually reveals three interconvertable isozymes [Ursprung and Leone; Johnson and Denniston; Grell et al., 1965; Ursprung and Carlin, 1968, Ann. N.Y. Acad. Sci. 151: 456-75; Jacobson, Murphy and Hartmann, 1970, J. Biol. Chem. 245: 1075-83; Jacobson and Pfuderer, 1970, J. Biol. Chem. 245: 3938-44; Jacobson, Murphy and Ortiz, 1972, Arch. Biochem. Biophys. 149: 22-35; Knopp and Jacobson, 1972, Arch. Biochem. Biophys. 149: 36-41; Schwartz, Gerace, O'Donnell and Sofer, 1975, Isoenzymes (C. Markert, ed.). Academic Press, Vol. I, pp. 725-51]. These vary in activity and stability, the most cathodal being more active, but less stable, than the more anodal forms. They probably result from the binding of 0, 1 or 2 moles per mole of a NAD+ addition complex with a carbonyl compound [Schwartz and Sofer, 1976, Nature (London) 263: 129-31; Schwartz, O'Donnell and Sofer, 1979, Arch. Biochem. Biophys. 194: 365-78; Winberg, Thatcher and McKinley-McKee, 1983, Biochem. Genet. 21: 63-80]. Feeding flies acetone, propan-2-ol, or 3-hydroxy-butanone, for example, converts isozymes to most anodal form and results in loss of enzyme activity in vitro and in vivo (Schwartz and Sofer, 1976; Papel, Henderson, van Herrewege, David and Sofer, 1979, Biochem. Genet. 17: 533-63). ADH has been purified (Sofer and Ursprung, 1968, J. Biol. Chem. 243: 3118-25; Schwartz et al., 1975; Thatcher, 1977, Biochem. J. 163: 317-23; Leigh Brown and Lee, 1979, Biochem. J. 179: 479-82; Juan and Gonzalez-Duarte, 1980, Biochem. J. 189: 105-10; Elliot and Knopp, 1975, Methods Enzymol. 41: 374-79; Chambers, 1984, Biochem. Genet. 22: 529-50). It is a homodimer with monomeric subunit molecular weight of 27500 daltons (Thatcher, 1980, Biochem. J. 187: 875-83); molecular extinction coefficient 4.8 X 104 liter/mol/cm (Juan and Gonzalez-Duarte, for ADH-S). Complete amino acid sequence determined by Thatcher (1980; see also Schwartz and Jornvall, 1976, Europ. J. Biochem. 68: 159-68; Auffret, Williams and Thatcher, 1978, FEBS Lett. 90: 324-26; Benyajati, Place, Powers, and Sofer, 1981, Proc. Nat. Acad. Sci. USA 78: 2317-21; Chambers, Laver, Campbell and Gibson, 1981, Proc. Nat. Acad. Sci. USA 78: 3103-07) with secondary structure predictions (Thatcher and Sawyer, 1980, Biochem J. 187: 884-86; Benyajati et al., 1981). Limited homology in supposed catalytic region with ribitol dehydrogenase of Klebsiella (Jornvall, Persson and Jeffry, 1981, Proc. Nat. Acad. Sci. USA 78: 4226-30). ADH shows a broad substrate specificity but is more active (by at least a factor of 5) with secondary than primary alcohols and shows highest activity to 3-6 carbon alcohols (Sofer and Ursprung; Thatcher and Camfield, 1977, Winberg et al., 1982, Chambers et al.). Differences in substrate specificity, kinetic constants and stability of different electrophoretic variants often reported (Anderson and McDonald, 1983, Proc. Nat. Acad. Sci. USA 80: 4798-802). Considerable heterogeneity in the specific activity of ADH within and between different AdhF and AdhS strains, though AdhS strains tend to be lower than AdhF [Day, Hillier and Clarke, 1974, Biochem. Genet. 11: 141-53, 155-65; Day and Needham, 1974, Biochem. Genet. 11: 167-75; Gibson, 1970, Nature (London) 227: 959-61; Gibson, Chambers, Wilkes and Oakeshott, 1980, Aust. J. Biol. Sci. 33: 479-89; Gibson and Miklovitch, 1971, Experientia 27: 99-100; Kreitman, 1980, Genetics 95: 467-75; Oakeshott, 1976, Aust. J. Biol. Sci. 29: 365-73; Sampsell, 1977, Biochem. Genet. 15: 971-88; Sampsell and Sims, 1982, Nature (London) 296: 853-55; Thorig, Schoone and Scharloo, 1975; Biochem. Genet. 13: 721-31; Vigue and Johnson; Hewitt, Pipkin, Williams and Chakrabartty, 1974, J. Hered. 65: 141-44; Ward, 1974, Biochem. Genet. 12: 449-58; Ward, 1975, Genet. Res. 26: 81-93; Maroni, Laurie-Ahlberg, Adams and Wilton, 1982, Genetics 101: 431-66; Rasmuson, Nilson and Zeppezauer, 1966, Hereditas 56: 313-16; Clarke, Camfield, Garvin and Pitts, 1979, Nature (London) 180: 517-18; Laurie-Ahlberg, Maroni, Bewley, Lucchesi and Weil, 1980, Proc. Nat. Acad. Sci. USA 77: 1073-77; Barnes and Birley, 1978, Heredity 40: 51-57; Barnes and Birley, 1978, Biochem. Genet. 16: 155-65; McDonald and Ayala, 1978, Genetics 89: 371-88; McDonald et al., 1980; Lewis and Gibson, 1978, Biochem. Genet. 16: 159-70]. With the exception of the studies by Thatcher and Sheik (1981, Biochem. J. 197: 111-17), Winberg et al. (1982), McDonald, Anderson and Santos (1980, Genetics 95: 1013-22); Eisses, Schoonen, Aben, Scharloo, and Thorig (1985, Mol. Gen. Genet. 199: 76-81) and Moxon et al. (1985), these were all done with crude extracts and not purified enzyme. Thatcher and Sheikh find the relative thermostabilities to be ADH-S > ADH-F > ADH-n5 > ADH-D. ADH-S shows slower dissociation of NADH from NADN-enzyme complex than ADH-F (Winberg, Hovik, and McKinley-McKee, 1985, Biochem. Genet. 23: 205-16). ADH is not a metalloenzyme (Place, Powers and Sofer, 1980, Fed. Proc. 39: 1640); but, paradoxically, is inhibited by certain metal ion chelators, e.g. pyrazole (Place, Powers and Sofer; Winberg et al., 1982; Moxon et al., 1985). Utilization of ethanol as an energy source (van Herrewege and David, 1974, C. Rend. Acad. Sci. Paris 279D: 335-38; van Herrewege, David and Grantham, 1980, Experientia 36: 846-47; Libion-Mannaert, Delcour, Deltombe-Lietaert, Lenelle-Montfort and Elens, 1976, Experentia 32: 22-23) depends on ADH activity (David, Bocquet, van Herrewege, Fouillet and Arens, 1978, Biochem. Genet. 16: 203-11). AdhF homozygotes usually show a better ability to survive on ethanol as a sole energy source than AdhS homozygotes (Daly and Clarke, 1981, Heredity 46: 219-26; Anderson, McDonald and Santos, 1981, Experientia 37: 463-64). AdhF and AdhS homozygotes also show behavioural differences in their response to ethanol (Parsons, 1977 Oecologia 30: 141-46; Cavener, 1979, Behav. Genet. 9: 359-65; Gelan and McDonald, 1980, Behav. Genet. 10: 237-49; Hougonto, Lietaert, Libion-Mannaert, Feytmans and Elens, 1982, Genetica 58: 121-28; Parsons, 1980, Behav. Genet. 10: 183-90; Parsons, 1980, Experientia 36: 1070-71). D. simulans enzyme monomers form heterodimers with those of D. melanogaster (E.H. Grell); D. simulans enzyme purified (Juan and Gonzalez-Duarte, 1981, Biochem. J. 195: 61-69). Sequence of D. simulans ADH (from DNA) similar to that of AdhS with following changes: ser1 -> ala1; gln82 -> lys82; ile184 -> val184 (Bodmer and Ashburner, 1984, Nature 309: 425-30). D. simulans and D. melanogaster enzymes differentially regulated in hybrids (Dickenson, Rowan, and Brennan, 1984, Heredity 52: 215-25). The Adh genes from D. orena and D. mauritiana have also been sequenced (Bodmer and Ashburner), and those of D. erecta, D. teissieri and D. yakuba mapped with restriction enzymes (Langley, Montgomery and Quattlebaum, 1982, Proc. Nat. Acad. Sci. USA 79: 5631-35).