Ethanol tolerance, alcohol dehydrogenase (ADH; EC 1.1.1.1) activity, and tissue-specific expression were examined in species of the cardini group of Drosophila using D. melanogaster as a standard of comparison. In contrast to most fruit-breeding species, all cardini species examined, two from the cardini subgroup and five from the dunni subgroup, were ethanol sensitive (LC50 < or = 2.05%) and the mean ADH activity of males ranges from only 8 to 16% that of D. melanogaster AdhFF. Among all seven cardini species, there were small but significant differences in ethanol tolerance and ADH activity. Differences in enzyme mobility were in accordance with the proposed phylogeny for the dunni-subgroup species. ADH is expressed in the fat body and midgut. Males of D. acutilabella and of D. belladunni have significantly less ethanol tolerance and express less ADH activity than females in zymograms and histological preparations.
We sequenced 2123 bp of the Adh-Adhr genomic region of Drosophila dunni of the cardini group from two cloned DNA PCR fragments and from two cDNA clones of an Adh transcript. This comprises the Adh coding region and introns, 3' UTR, intergenic sequence, and most of Adhr, which is 260 bp downstream of Adh. Both genes have the typical Drosophila melanogaster Adh structure of three exons and two introns, except for changes in the putative 8 bp sequence involved in downregulation within the 3' UTR of Adh. Two amino acid substitutions could explain the low activity previously reported for this enzyme in D. dunni: Thr --> Lys at position 191 and Val --> Thr at position 189. D. dunni's Adh has the lowest codon bias reported so far for Drosophila species, and based on analysis of the nucleotide substitution rate, it is less conserved than Adhr.
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