scholarly journals Identification of cis-regulatory elements required for larval expression of the Drosophila melanogaster alcohol dehydrogenase gene.

Genetics ◽  
1990 ◽  
Vol 124 (3) ◽  
pp. 637-646 ◽  
Author(s):  
V Corbin ◽  
T Maniatis
Keyword(s):  
Drosophila Melanogaster ◽  
Alcohol Dehydrogenase ◽  
Dna Sequences ◽  
Regulatory Elements ◽  
Malpighian Tubules ◽  
Proximal Promoter ◽  
Start Site ◽  
Wild Type ◽  
Tissue Specific ◽  
Adh Gene

Abstract The Alcohol dehydrogenase (Adh) genes of two distantly related species, Drosophila melanogaster and Drosophila mulleri, display similar, but not identical, patterns of tissue-specific expression in larvae and adults. The regulatory DNA sequences necessary for wild-type Adh expression in D. mulleri larvae were previously reported. In this paper we present an analysis of the DNA sequences necessary for wild-type Adh expression in D. melanogaster larvae. We show that transcription from the proximal promoter of the melanogaster Adh gene is regulated by a far upstream enhancer and two or more elements near the transcription start site. The enhancer is tissue specific and stimulates transcription to high levels in fat body and to lower levels in midgut and malpighian tubules whether linked to the proximal promoter or to a heterologous promoter. The enhancer activity localized to at least two discrete regions dispersed over more than 1.7 kb of DNA. Deletion of any one of these subregions reduces Adh transcription in all three larval tissues. Similarly, two regions immediately upstream of the proximal promoter start site are necessary for wild-type transcription levels in all three tissues. Thus, each of the identified regulatory elements is sufficient for low levels of Adh gene expression in all three larval tissues, but maximal levels of expression requires the entire set.

scholarly journals Insertion of a copia element 5' to the Drosophila melanogaster alcohol dehydrogenase gene (adh) is associated with altered developmental and tissue-specific patterns of expression.

Genetics ◽  
1989 ◽  
Vol 121 (4) ◽  
pp. 787-794
Author(s):  
D J Strand ◽  
J F McDonald
Keyword(s):  
Drosophila Melanogaster ◽  
Alcohol Dehydrogenase ◽  
Transcriptional Start Site ◽  
Adult Life ◽  
Life Stages ◽  
Start Site ◽  
Wild Type ◽  
Dehydrogenase Gene ◽  
In Trans ◽  
Copia Element

Abstract The Drosophila melanogaster alcohol dehydrogenase gene (adh) is under the control of two separate promoters (proximal and distal) which are preferentially utilized at the larval and adult life stages, respectively. A variant alcohol dehydrogenase allele (RI-42) isolated from a natural population contains a copia retroviral-like transposable element inserted 240 bp upstream from the distal (adult) adh transcriptional start site. Levels of adh transcripts in the RI-42 variant are reduced in tissues and at life stages where copia is actively expressed and are affected in trans- by mutant alleles at the suppressor-of-white-apricot (su(wa] and suppressor-of-forked (su(f] loci. These suppressor genes have no effect on adh expression in wild-type Drosophila.

scholarly journals Analysis of sequences regulating larval expression of the Adh gene of Drosophila melanogaster.

Genetics ◽  
1991 ◽  
Vol 129 (3) ◽  
pp. 763-771
Author(s):  
N L Shen ◽  
E C Hotaling ◽  
G Subrahmanyam ◽  
P F Martin ◽  
W Sofer
Keyword(s):  
Enzyme Activity ◽  
Drosophila Melanogaster ◽  
Alcohol Dehydrogenase ◽  
Reference Gene ◽  
Base Pair ◽  
Regulatory Region ◽  
Regulatory Elements ◽  
Base Pair Deletion ◽  
Adh Gene ◽  
Tata Sequence

Abstract The effects of a series of eight, 50 base pair internal deletions in the 5' region upstream of the proximal transcription start site of the Adh gene of Drosophila melanogaster were examined in a quantitative assay. Mixtures of two plasmids, one bearing a deleted gene, the other with an intact reference gene, were injected into alcohol dehydrogenase-negative embryos. Third instar larvae of the injected generation were assayed for relative alcohol dehydrogenase enzyme activity. Quantitative analysis of the eight deletions indicated that two regions were required for any detectable enzyme activity and one region was required for appropriate tissue specificity. The remaining five deletions significantly decreased, but did not eliminate activity. When the deleted genes were placed on a plasmid with an intact reference gene, activities of all but one deletion were restored to levels equivalent to that of the intact reference gene (regardless of orientation). This restoration of activity did not occur when the regulatory region of the intact gene was replaced with the Hsp70 heat shock promoter nor when the 50-base pair deletion encompassed the region that includes the TATA sequence. The fact that seven of the eight deleted genes express activity in the presence of a reference gene on the same plasmid suggests that the deleted gene is controlled by regulatory elements in the reference gene. Further, these regulatory elements exhibit no preference for their own, more proximate, promoter.

scholarly journals Molecular control of the induction of alcohol dehydrogenase by ethanol in Drosophila melanogaster larvae.

Genetics ◽  
1990 ◽  
Vol 124 (4) ◽  
pp. 881-888 ◽  
Author(s):  
A M Kapoun ◽  
B W Geer ◽  
P W Heinstra ◽  
V Corbin ◽  
S W McKechnie
Keyword(s):  
Drosophila Melanogaster ◽  
Alcohol Dehydrogenase ◽  
Dna Sequence ◽  
P Element ◽  
Start Site ◽  
Major Pathway ◽  
Molecular Control ◽  
Adh Gene ◽  
Regulatory Dna ◽  
Transcript Start Site

Abstract The activity of alcohol dehydrogenase (ADH:EC 1.1.1.1), the initial enzyme in the major pathway for ethanol degradation, is induced in Drosophila melanogaster larvae by low concentrations of dietary ethanol. Two lines of evidence indicate that the metabolic products of the ADH pathway for ethanol degradation are not directly involved in the induction of Adh. First, the accumulation of the proximal transcript in Adhn2 larvae was increased when the intracellular level of ethanol was elevated. In addition, the ADH activity, the proximal Adh mRNA, and the intracellular concentration of ethanol were elevated coordinately in wild-type larvae fed hexadeuterated-ethanol, which is metabolized more slowly than normal ethanol. An examination of P element transformant lines with specific deletions in the 5' regulatory DNA of the Adh gene showed that a DNA sequence between +527 and +604 of the distal transcript start site is essential for the induction of the Adh gene [corrected]. The DNA sequence between -660 and about -5000 of the distal transcript start site was important for the down-regulation of the induction response.

Die fluoreszierenden Stoffe aus den Malpighischen-Gefäßen der Wildform und verschiedener Augenfarbenmutanten von Drosophila melanogaster

1968 ◽  
Vol 23 (3) ◽  
pp. 376-386 ◽  
Author(s):  
Armin Wessing ◽  
Dieter Eichelberg
Keyword(s):  
Drosophila Melanogaster ◽  
Malpighian Tubules ◽  
Wild Type ◽  
Eye Color ◽  
Eye Color Mutants

The Malpighian tubules of Drosophila melanogaster accumulate a great number of substances, many of which fluoresce. This paper is concerned with the identification of these substances by chromatography and their location by fluorescentmicroscopy (fig. 4, 5). It appears that they mainly belong to the following three groups: Pteridines, tryptophane and some of its metabolites, and riboflavine (tab. 1).The pattern of fluorescent substances of the eye color mutants cn, v, se, st, bw, ry, and w vary significantly. The patterns of these mutants are compared and discussed with that of the wild-type.

Molecular analysis of alcohol dehydrogenase electromorphs in wild type and transformed Drosophila melanogaster

1988 ◽  
Vol 150 (2) ◽  
pp. 655-664 ◽  
Author(s):  
Mark A. Batzer ◽  
Trent D. Desselle ◽  
Mark D. Brennan ◽  
William R. Lee ◽  
Bruce Tedeschi
Keyword(s):  
Drosophila Melanogaster ◽  
Alcohol Dehydrogenase ◽  
Molecular Analysis ◽  
Wild Type

scholarly journals Tissue-specific expression of the diazepam-binding inhibitor in Drosophila melanogaster: cloning, structure, and localization of the gene.

1994 ◽  
Vol 14 (10) ◽  
pp. 6983-6995 ◽  
Author(s):  
M Kolmer ◽  
C Roos ◽  
M Tirronen ◽  
S Myöhänen ◽  
H Alho
Keyword(s):  
Drosophila Melanogaster ◽  
Transcription Initiation ◽  
Coenzyme A ◽  
Cpg Island ◽  
High Energy ◽  
Malpighian Tubules ◽  
Rnase Protection ◽  
Tissue Specific ◽  
Receptor Modulation ◽  
Acyl Coenzyme A

The diazepam-binding inhibitor (DBI; also called acyl coenzyme A-binding protein or endozepine) is a 10-kDa polypeptide found in organisms ranging from yeasts to mammals. It has been shown that DBI and its processing products are involved in various specific biological processes such as GABAA/benzodiazepine receptor modulation, acyl coenzyme A metabolism, steroidogenesis, and insulin secretion. We have cloned and sequenced the Drosophila melanogaster gene and cDNA encoding DBI. The Drosophila DBI gene encodes a protein of 86 amino acids that shows 51 to 56% identity with previously known DBI proteins. The gene is composed of one noncoding 5' and two coding exons and is localized on the chromosomal map at position 65E. Several transcription initiation sites were detected by RNase protection and primer extension experiments. Computer analysis of the promoter region revealed features typical of housekeeping genes, such as the lack of TATA and CCAAT elements. However, in its low GC content and lack of a CpG island, the region resembles promoters of tissue-specific genes. Northern (RNA) analysis revealed that the expression of the DBI gene occurred from the larval stage onwards throughout the adult stage. In adult flies, DBI mRNA and immunoreactivity were detected in the cardia, part of the Malpighian tubules, the fat body, and gametes of both sexes. Developmentally regulated expression, disappearing during metamorphosis, was detected in the larval and pupal brains. No expression was detected in the adult nervous system. On the basis of the expression of DBI in some but not all tissues with high energy consumption, we propose that in D. melanogaster, DBI is involved in energy metabolism in a manner that depends on the substrate used for energy production.

scholarly journals Regulation of enzyme activities in Drosophila: I. The detection of regulatory loci by gene dosage responses

1974 ◽  
Vol 24 (1) ◽  
pp. 59-72 ◽  
Author(s):  
John M. Rawls ◽  
John C. Lucchesi
Keyword(s):  
Enzyme Activity ◽  
Drosophila Melanogaster ◽  
Alcohol Dehydrogenase ◽  
Gene Dosage ◽  
Regulatory Elements ◽  
Gpdh Activity ◽  
Regulatory Loci ◽  
Chromosome Ii ◽  
Chromosome Segments ◽  
Regulatory Sites

SUMMARYIn order to detect regulatory genetic sites in the autosomes of Drosophila melanogaster, the levels of X-linked glucose-6-phosphate dehydro-genase and autosomally linked α-glycerophosphate and isocitrate dehydrogenases have been monitored in extracts of flies aneuploid for regions of chromosomes II and III. In addition to expected structural gene dosage responses of α-GPDH and IDH, flies hyperploid for several autosome regions were found to display altered levels of one or more of the enzymes studied. While IDH activity was increased in flies hyperploid for segments of both chromosomes II and III, α-GPDH activity was decreased in specific hyperploids for chromosome II regions only. The latter group of segmental aneuploids were normal with respect to levels of chromosome II-linked alcohol dehydrogenase. To test if the observed responses were due to dosage changes of discrete genes lying within the larger effective segments, flies aneuploid for subdivisions of the chromosome segments 21A-25CD, 35A–40, and 70CD–71B were assayed. For two of these large segments so analysed, the apparent effects were attributable to specific small subdivisions, suggesting the presence of discrete regulatory sites within the latter. For the 35A–40 region the α-GPDH effect observed for subdivisions was not sufficient to account for the large α-GPDH decrease seen in flies hyperploid for the large, inclusive region. These observations are discussed with respect to the possible bases of effect of regulatory elements on enzyme activity.

scholarly journals Molecular cloning of suppressor of sable, a Drosophila melanogaster transposon-mediated suppressor.

1986 ◽  
Vol 6 (5) ◽  
pp. 1520-1528 ◽  
Author(s):  
D Y Chang ◽  
B Wisely ◽  
S M Huang ◽  
R A Voelker
Keyword(s):  
Drosophila Melanogaster ◽  
Dna Sequences ◽  
Insertion Site ◽  
Hybrid Dysgenesis ◽  
P Element ◽  
Dna Transformation ◽  
Wild Type ◽  
Induced Mutations ◽  
X Ray ◽  
Element Insertion

A hybrid dysgenesis-induced allele [su(s)w20] associated with a P-element insertion was used to clone sequences from the su(s) region of Drosophila melanogaster by means of the transposon-tagging technique. Cloned sequences were used to probe restriction enzyme-digested DNAs from 22 other su(s) mutations. None of three X-ray-induced or six ethyl methanesulfonate-induced su(s) mutations possessed detectable variation. Seven spontaneous, four hybrid dysgenesis-induced, and two DNA transformation-induced mutations were associated with insertions within 2.0 kilobases (kb) of the su(s)w20 P-element insertion site. When the region of DNA that included the mutational insertions was used to probe poly(A)+ RNAs, a 5-kb message was detected in wild-type RNA that was present in greatly reduced amounts in two su(s) mutations. By using strand-specific probes, the direction of transcription of the 5-kb message was determined. The mutational insertions lie in DNA sequences near the 5' end of the 5-kb message. Three of the seven spontaneous su(s) mutations are associated with gypsy insertions, but they are not suppressible by su(Hw).

Tissue-Specific Regulatory Elements in the Human Alcohol Dehydrogenase 6 Gene

2000 ◽  
Vol 19 (8) ◽  
pp. 487-497 ◽  
Author(s):  
Xin Zhi ◽  
Edward M. Chan ◽  
Howard J. Edenberg
Keyword(s):  
Alcohol Dehydrogenase ◽  
Regulatory Elements ◽  
Tissue Specific

scholarly journals The organization and expression of the light gene, a heterochromatic gene of Drosophila melanogaster.

Genetics ◽  
1990 ◽  
Vol 125 (1) ◽  
pp. 129-140 ◽  
Author(s):  
R H Devlin ◽  
B Bingham ◽  
B T Wakimoto
Keyword(s):  
Drosophila Melanogaster ◽  
Dna Sequences ◽  
Molecular Mapping ◽  
Single Copy ◽  
Transcription Unit ◽  
Malpighian Tubules ◽  
Centromeric Heterochromatin ◽  
Chromosome 2 ◽  
Induced Mutations ◽  
Flanking Regions

Abstract The light (lt) gene is located in the centromeric heterochromatin of chromosome 2 of Drosophila melanogaster. This gene is necessary for normal levels of pigmentation in a number of adult and larval tissues and is required for viability. Hybrid dysgenic and X-ray induced mutations have been used to identify the gene and compare its organization to that of euchromatic genes. Molecular mapping of lt mutations and its major transcripts has shown that the lt gene is at least 17 kb. By injecting cosmid clones that include this region into lt mutant embryos, we have defined a 30-kb region that can transiently rescue the pigmentation defect in the Malpighian tubules. The major transcription unit of this gene is comprised of exons that are single copy. It is unusual in its organization in having a heterogeneous array of middle repetitive DNA sequences within its intronic and flanking regions.

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