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D A Hickey

Publications and source records attributed to D A Hickey.

At least 37 records · Page 2Linked to original sources
PubMed1990

Amylase gene expression in intraspecific and interspecific somatic transformants of Drosophila.

The Amylase locus in Drosophila melanogaster normally contains two copies of the structural gene for alpha-amylase, a centromere-proximal copy, Amy-p, and a distal copy, Amy-d. Products of the two genes may display discrete electrophoretic mobilities, but many strains known to carry the Amy duplication are characterized by a single amylase electromorph, e.g., Oregon-R, which produces the mobility variant AMY-1. A transient expression assay was used in somatic transformation experiments to test the functional status of the Amy genes from an Oregon-R strain. Plasmid constructs containing either the proximal or distal copy were tested in amylase-null hosts. Both genes produced a functional AMY-1 isozyme. Constructs were tested against an AMY-3 reference activity produced by a coinjected plasmid that contains the Amy-d3 allele from a Canton-S strain. With reference to the internal control, the Amy-p and Amy-d genes from Oregon-R expressed different relative activity levels for AMY-1 in transient assays. The transient expression assay was successfully used to test the functional status of Amy-homologous sequences from strains of other species of Drosophila characterized by a single amylase elctromorph, namely, Drosophila pseudoobscura ST and Drosophila miranda S 204. The amylase-null strain of D. melanogaster provided the hosts for these interspecific somatic transformation experiments.

Amylases↗
PubMed1989

A general model for the evolution of nuclear pre-mRNA introns.

We present an overview of the evolution of eukaryotic split gene structure and pre-mRNA splicing mechanisms. We have drawn together several seemingly conflicting ideas and we show that they can all be incorporated in a single unified theory of intron evolution. The resulting model is consistent with the notion that introns, as a class, are very ancient, having originated in the "RNA world"; it also supports the concept that introns may have played a crucial role in the construction of many eukaryotic genes and it accommodates the idea that introns are related to mobile insertion elements. Our conclusion is that introns could have a profound effect on the course of eukaryotic gene evolution, but that the origin and maintenance of intron sequences depends, largely, on natural selection acting on the intron sequences themselves.

Biological Evolution↗
PubMed1989

Molecular biology of enzyme adaptations in higher eukaryotes.

Multicellular eukaryotes have evolved complex homeostatic mechanisms that buffer the majority of their cells from direct interaction with the external environment. Thus, in these organisms long-term adaptations are generally achieved by modulating the developmental profile and tissue specificity of gene expression. Nevertheless, a subset of eukaryotic genes are still involved in direct responses to environmental fluctuations. It is the adaptative responses in the expression of these genes that buffers many other genes from direct environmental effects. Both microevolutionary and macroevolutionary patterns of change in the structure and regulation of such genes are illustrated by the sequences encoding alpha-amylases. The molecular biology and evolution of alpha-amylases in Drosophila and other higher eukaryotes are presented. The amylase system illustrates the effects of both long-term and short-term natural selection, acting on both the structural and regulatory components of a gene--enzyme system. This system offers an opportunity for linking evolutionary genetics to molecular biology, and it allows us to explore the relationship between short-term microevolutionary changes and long-term adaptations.

Adaptation, Biological↗
PubMed1988

DNA rearrangement causes multiple changes in gene expression at the amylase locus in Drosophila melanogaster.

A spontaneous null mutation at the alpha-amylase locus in Drosophila melanogaster was recovered from a laboratory population. The mutant strain was found to lack amylase enzyme production and to produce low, but detectable, levels of amylase mRNA. Moreover, the null strain is also lacking the glucose repression of amylase mRNA production which is seen in wild-type strains. The mutant phenotype correlates with a rearrangement in genomic DNA which, in turn, corresponds to a simple inversion in the arrangement observed most frequently in North American populations of D. melanogaster, including the common laboratory strain, Oregon-R. These results have implications for our understanding of both the evolution of the duplicated amylase gene structure and the regulation of amylase gene expression.

Amylases↗
PubMed1988

Maintenance of the 2 micron circle plasmid of Saccharomyces cerevisiae by sexual transmission: an example of a selfish DNA.

Many eukaryotic mobile elements have been identified, but few have any obvious function. This has led to the proposal that many such elements may be parasitic DNA. We have used the 2 micron circle plasmid of Saccharomyces cerevisiae as a model system to investigate the maintenance of a cryptic genetic element. We find that under certain conditions this plasmid can spread through experimental populations despite demonstrable selection against it. This spread is dependent upon outbreeding, suggesting that cell to cell transmission of the plasmid during the yeast sexual cycle can counterbalance selection, and maintain the plasmid in populations. This result provides experimental support for the idea that some mobile elements may be parasitic DNA.

Crosses, Genetic↗
PubMed1988

Evolutionary conservation of the chromosomal configuration and regulation of amylase genes among eight species of the Drosophila melanogaster species subgroup.

Nuclear DNA was extracted from each of the eight species comprising the Drosophila melanogaster species subgroup. Southern hybridization of this DNA by using a molecular probe specific for the alpha-amylase coding region showed that the duplicated structure of the amylase locus, first found in D. melanogaster, is conserved among all species of the melanogaster subgroup. Evidence is also presented for the concerted evolution of the duplicated genes within each species. In addition, it is shown that the glucose repression of amylase gene expression, which has been extensively studied in D. melanogaster, is not confined to this species but occurs in all eight members of the species subgroup. Thus, both the duplicated gene structure and the glucose repression of Drosophila amylase gene activity are stable over extended periods of evolutionary time.

Amylases↗
PubMed1987

Enzyme-coding genes as molecular clocks: the molecular evolution of animal alpha-amylases.

We constructed a cDNA library for the beetle, Tribolium castaneum. This library was screened using a cloned amylase gene from Drosophila melanogaster as a molecular probe. Beetle amylase cDNA clones were isolated from this bank, and the nucleotide sequence was obtained for a cDNA clone with a coding capacity for 228 amino acids. Both the nucleotide sequence and predicted amino acid sequence were compared to our recent results for D. melanogaster alpha-amylases, along with published sequences for other alpha-amylases. The results show that animal alpha-amylases are highly conserved over their entire length. A broader comparison, which includes plant and microbial alpha-amylase sequences, indicates that parts of the gene are conserved between prokaryotes, plants, and animals. We discuss the potential importance of this and other enzyme-coding genes for the construction of molecular phylogenies and for the study of the general question of molecular clocks in evolution.

Amino Acid Sequence↗
PubMed1987

A Drosophila gene is subject to glucose repression.

Amylase-specific cDNA probes were used to assay amylase mRNA levels in third-instar larvae of Drosophila melanogaster. It is shown that there is a difference, of the order of 100-fold, in the mRNA levels between larvae that are fed 10% glucose and larvae of the same wild-type strain that are fed an equivalent diet lacking glucose. In fact, the glucose-fed larvae have barely detectable levels of amylase mRNA. This large difference in transcript abundance indicates that the glucose effect, which we previously characterized at the level of enzyme activity, probably reflects a change in the transcriptional activity of the amylase gene.

Journal Article↗
PubMed1987

Molecular cloning of DNA complementary to Drosophila melanogaster alpha-amylase mRNA.

Several lambda clones containing cDNAs from Drosophila melanogaster were identified in a lambda cDNA bank using two different approaches: (i) cross-species hybridization using a mouse amylase cDNA probe, and (ii) probing with a differential probe, generated from Drosophila RNA. An amylase cDNA fragment was used, in turn, for the isolation and characterization of amylase genomic clones. The size of the Drosophila amylase mRNA was estimated at 1650 b. This is comparable with the size of the murine amylase messenger that encodes a protein of similar molecular weight. In Drosophila larvae, amylase mRNA can account for as little as 0.01% of the poly(A)+ RNA under conditions of dietary glucose repression or greater than 1% of poly(A)+ RNA under derepressing dietary conditions.

Animals↗
PubMed1986

The alpha-amylase gene in Drosophila melanogaster: nucleotide sequence, gene structure and expression motifs.

We present the complete nucleotide sequence of a Drosophila alpha-amylase gene and its flanking regions, as determined by cDNA and genomic sequence analysis. This gene, unlike its mammalian counterparts, contains no introns. Nevertheless the insect and mammalian genes share extensive nucleotide similarity and the insect protein contains the four amino acid sequence blocks common to all alpha-amylases. In Drosophila melanogaster, there are two closely-linked copies of the alpha-amylase gene and they are divergently transcribed. In the 5'-regions of the two gene-copies we find high sequence divergence, yet the typical eukaryotic gene expression motifs have been maintained. The 5'-terminus of the alpha-amylase mRNA, as determined by primer extension analysis, maps to a characteristic Drosophila sequence motif. Additional conserved elements upstream of both genes may also be involved in amylase gene expression which is known to be under complex controls that include glucose repression.

Amino Acid Sequence↗
PubMed1986

Introns as relict retrotransposons: implications for the evolutionary origin of eukaryotic mRNA splicing mechanisms.

A model is presented for the evolutionary origin of intron sequences within eukaryotic protein-coding genes. We propose that introns are the vestiges of transposable elements and, specifically, that they represent a novel class of retrovirus-like transposons. The attraction of the retrotransposon model is that it gives the RNA splicing mechanism a central role in the evolution of introns. There is a growing body of evidence to suggest that several aspects of splicing are intron-encoded. Consequently, it is reasonable to look for evolutionary explanations of the splicing mechanism in the context of the evolution of the intron sequences themselves. According to this model the ancestral intron genomes were replicated into RNA copies simply because of their insertion within transcriptionally active regions of the host genome. Splicing was necessary not only to minimize their negative effects on host gene expression, but also, and perhaps more importantly, to generate new copies of the intron genome free of flanking exon sequences. These spliced intron copies were then available for reverse transcription and reinsertion elsewhere in the genome. Thus, splicing can be seen as an essential step in the intron replication cycle. Most modern introns have probably lost the majority of their original genetic content and may be considered as degenerate evolutionary relicts. An exception to this degeneracy is the set of splicing signals which must be retained because of its continued importance to host cell survival.(ABSTRACT TRUNCATED AT 250 WORDS)

Biological Evolution↗
PubMed1982

A molecular explanation of frequency-dependent selection in Drosophila.

Frequency-dependent selection provides a means for maintaining genetic variability within populations, without incurring a large genetic load. There is a wealth of experimental evidence for the existence of frequency-dependent changes in genotypic fitness among a wide variety of organisms. Examples of traits which have been shown to be subject to frequency-dependent selection include the self-incompatibility alleles of plants, chromosomal rearrangements in Drosophila, visible mutations, enzyme variants and rare-male mating advantage in Drosophila. These experiments have been interpreted in a number of different ways. Principally, frequency dependence of genotype fitness may result from intergenotype facilitation due to the production of biotic residues, or from the differential use of resources by the competing genotypes. However, it has proved extremely difficult to isolate and identify any biotic residue of importance or, alternatively, to understand the manner in which genotypes partition the environment. Thus, the difficulty in the interpretation of experiments which show frequency-dependent selective effects stems largely from our lack of understanding of the exact physiological mechanisms which produce these frequency-dependent effects. The principal aim of this study was to investigate the mechanisms associated with frequency-dependent selection at the amylase locus in Drosophila melanogaster. The excretion of catalytically active amylase enzyme and its effect on food medium composition were correlated with the outcome of intraspecific competition between amylase-deficient and amylase-producing genotypes. Amylase-producing genotypes were shown to excrete enzymatically active amylase protein into the food medium. The excreted amylase causes the external digestion of dietary starch; this accounts for the frequency-dependent increase in the viability of the amylase-deficient mutants in mixed cultures, maintained on a starch-rich diet.

Amylases↗
PubMed1982

Regulation of amylase activity in drosophila melanogaster: effects of dietary carbohydrate.

The level of amylase activity in larvae and adults of Drosophila melanogaster is dependent on the dietary carbohydrate source; flies or larvae from a food medium containing starch show higher levels of activity than individuals from a food containing simple sugars. This is shown to be due to repression of activity by sugars rather than enhancement of activity by starch. Moreover, the changes in enzyme activity reflect a change in enzyme quantity rather than a change in catalytic efficiency. The seeming stimulation of amylase activity by sucrose in some experiments is due, simply, to comparisons with "starvation" diets which cause a large nonspecific reduction in enzyme activity. Though all strains tested showed repression of enzyme activity by simple sugars, the degree of repression varies between strains. Also, in those strains which carry a duplication of the amylase structural gene, the two isozymal forms of amylase can be differentially repressed by dietary sugars.

Amylases↗
PubMed1981

Regulation of amylase activity in Drosophila melanogaster: variation in the number of enzyme molecules produced by different amylase genotypes.

Purified amylases from high- and low-activity variants of Drosophila melanogaster showed identical specific activities. Immunoelectrophoresis of crude larval homogenates showed severalfold differences between strains in the amounts of cross-reacting material. Control of amylase activity is "trans"-acting in heterozygotes between high- and low-activity variants. These results suggest the existence of polymorphic regulatory genes affecting the production levels of amylase protein in D. melanogaster.

Amylases↗
PubMedDate not supplied

Selfish DNA: a sexually-transmitted nuclear parasite.

A quantitative population genetics model for the evolution of transposable genetic elements is developed. This model shows that "selfish" DNA sequences do not have to be selectively neutral at the organismic level; indeed, such DNA can produce major deleterious effects in the host organism and still spread through the population. The model can be used to explain the evolution of introns within eukaryotic genes; this explanation does not invoke a long-term evolutionary advantage for introns, nor does it depend on the hypothesis that eukaryotic gene structure may be an evolutionary relic. Transposable genes that carried information specifying sexual reproduction in the host organism would favor their own spread. Consequently, it is tempting to speculate that some of the genes controlling sex were originally selected as transposable elements.

Base Sequence↗