Physiological Significance of the Alcohol Dehydrogenase Polymorphism in Larvae of Drosophila

This study deals with biochemical and metabolic-physiological aspects of the relationship between variation in in vivo alcohol dehydrogenase activity and fitness in larvae homozygous for the alleles Adh71k, AdhF, AdhS, of Drosophila melanogaster, and for the common Adh allele of Drosophila simulans. The Adh genotypes differ in the maximum oxidation rates of propan-2-ol into acetone in vivo. There are smaller differences between the Adh genotypes in rates of ethanol elimination. Rates of accumulation of ethanol in vivo are negatively associated with larval-to-adult survival of the Adh genotypes. The rank order of the maximum rates of the ADHs in elimination of propan-2-ol, as well as ethanol, is ADH-71k greater than ADH-F greater than ADH-S greater than simulans-ADH. The ratio of this maximum rate to ADH quantity reveals the rank order of ADH-S greater than ADH-F greater than ADH-71k greater than simulans-ADH, suggesting a compensation for allozymic efficiency by the ADH quantity in D. melanogaster.Our findings show that natural selection may act on the Adh polymorphism in larvae via differences in rates of alcohol metabolism.     

The Effects of Disruptive and Stabilizing Selection on Body Size in DROSOPHILA MELANOGASTER. II. Analysis of Responses in the Thorax Selection Lines

An analysis was made of changes in mean and variance in some thorax selection lines. The decrease of mean thorax length in the stabilizing selection lines (S) was a consequence of a directional selection component, caused by the skewness of the frequency distributions. The slight or temporary increase of the phenotypic variance and the large increase of the mean value in the disruptive selection lines with random mating (D(R)) could be attributed to differences in reproduction between small and large flies (egg production and mating success). Phenotypic variability was high in two disruptive selection lines with compulsory mating of opposite extremes (D(-)). The mechanism of the change in variability was different in these replicate lines. In D(-)-1 the change was obtained by an increase of the environmental and the nonadditive genetic components of the variance. In D(-)-2 almost exclusively an increase of additive genetic variance occurred.     

Genetic Localization and Action of Regulatory Genes and Elements for Tissue-Specific Expression of alpha-Amylase in DROSOPHILA MELANOGASTER

Regulation of tissue-specific alpha-amylase (Amy) expression in Drosophila melanogaster was investigated with a newly developed method that detects the distribution of alpha-amylase allozymes in midguts of single adults or third-instar larvae. Trans regulation was found for alpha-amylase production in the posterior midgut (PMG) of adults, whereas cis regulation was demonstrated for the larval midgut. Independent regulation of components of the duplicated Amy locus was found in larvae. Recombination between the components of the Amy locus did not result in separation of the putative, very closely linked (less than 0.1 cM) cis-acting regulatory elements for alpha-amylase expression in the anterior midgut (AMG) of larvae. The expression of one of the components of the duplicated Amy locus in the AMG of larvae was influenced by a regulatory gene that was mapped at 2-79.1. alpha-Amylase expression in the adult PMG was controlled by a trans-acting regulatory gene localized at 2-79.0, in agreement with the data of Abraham and Doane.     

Functional significance of amylase polymorphism in Drosophila melanogaster. III. Ontogeny of amylase and some α-glucosidases

Changes in amylase (E.C. 3.2.1.1), maltase (E.C. 3.2.1.20), sucrase, and PNPGase activities in relation to changes in wet weight and protein content were studied during the development of larvae and adult flies from two strains of Drosophila melanogaster, homozygous for different amylase alleles. All -glucosidase activities increase exponentially during a large part of larval development, parallel to the increase in weight, and drop at the end of the third instar. Amylase activity of the Amy ¹ strain follows the same pattern. In contrast, amylase activity of the Amy ^4,6 strain continues its exponential increase longer. In the third larval instar amylase activity in the Amy ^4,6 strain becomes much higher than in the Amy ¹ strain. During the first hours of adult life amylase activity of the two strains does not differ. Then Amy ^4,6 activity starts to rise and becomes much higher (4–5 times) than Amy ¹ amylase activity, which remains approximately constant. All adult enzyme activities are much higher than in larvae. Comparison of enzyme activity of amylase and -glucosidases in larvae and adults confirms that differences in amylase activities can become important only when starch is a limiting factor in the food.The investigations were supported by the Foundation for Fundamental Biological Research (BION), which is subsidized by the Netherlands Organization for the Advancement of Pure Research (Z.W.O.).     

Kinetics and thermodynamics of ethanol oxidation catalyzed by genetic variants of the alcohol dehydrogenase from Drosophila melanogaster and D. simulans

Four naturally occurring variants of the alcohol dehydrogenase enzyme (ADH; EC 1.1.1.1) from Drosophila melanogaster and D. simulans, with different primary structures, have been subjected to kinetic studies of ethanol oxidation at five temperatures. Two amino acid replacements in the N-terminal region which distinguish the ADH of D. simulans from the three ADH allozymes of D. melanogaster generate a significantly different activation enthalpy and entropy, and Gibbs free energy change. The one or two amino acid replacements in the C-terminal region between the ADH allozymes of D. melanogaster do not have such clear-cut effects. All four ADH variants show highly negative activation entropies. Sarcosine oxidation by the ADH-71k variant of D. melanogaster has an activation energy barrier similar to that of ethanol oxidation. Three amino acid differences between the ADH of D. simulans and the ADH-F variant of D. melanogaster influence the kappa cat and kappa cat/Kethm constant by a maximum factor of about 2 and 2.5, respectively, over the whole temperature range. Product inhibition patterns suggest a 'rapid equilibrium random' mechanism of ethanol oxidation by the ADH-71k, and the ADH of D. simulans.     

Variation between electrophoretically identical alleles at the alcohol dehydrogenase locus in Drosophila melanogaster

A new variant of alcohol dehydrogenase (ADH 7lk) was found in a laboratory stock of Drosophila melanogaster. ADH in this stock had the same electrophoretic mobility as the F variant both on acrylamide and on agar. Activity levels were similar to the levels in F flies at temperature between 15 and 25 C. But while ADH F enzyme is inactivated rapidly at 40 C, ADH 7lk is still active. Also, ADH S is not inactivated at this temperature, but has a far lower activity per fly than ADH 7lk. Genetic analysis showed that the new variant is an allele of the Adh locus.     

The functional significance of amylase polymorphism in Drosophila melanogaster VI. Duration of development and amylase activity in larvae when starch is a limiting factor

Two stocks homozygous for the amylase alleles Amy ¹ and Amy ^{4, 6} were compared for amylase activity during larval development and duration of larval development on a food medium on which addition of starch promotes survival. The two stocks have the same development time when reared under optimal food conditions. Then, the higher amylase activity which characterizes the Amy ^{4, 6} stock appears far into the third instar. On the food used in the present experiments in which starch is a limiting factor for survival, the activity difference appears already in the 2nd instar. Moreover, Amy ^{4, 6} has a duration of larval development which is almost one day shorter than the Amy ¹ stock. This would result in a large selective advantage for the Amy ^{4, 6} stock when brought in competition on this food with the Amy ¹ stock.These rrsults support the explanation given for the results of earlier competition experiments between the Amy ¹ and the Amy ^{4, 6} stock, that the relative fitness of the stock with the high amylase activity improves when starch is added to a food medium in which it is a limiting factor for survival because the Amy ^{4, 6} stock by virtue of its higher amalyse activity utilizes starch better than its competitor.