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Title: The alcohol dehydrogenase gene is nested in the outspread locus of Drosophila melanogaster

Abstract

This report describes the structure and expression of the outspread (osp) gene of Drosophila melanogaster. Previous work showed that chromosomal breakpoints associated with mutations of the osp locus map to both sides of the alcohol dehydrogenase gene (Adh), suggesting that Adh and the adjacent gene Adh{sup r} are nested in osp. We extended a chromosomal walk and mapped additional osp mutations to define the maximum molecular limit of osp as 119 kb. We identified a 6-kb transcript that hybridizes to osp region DNA and is altered or absent in osp mutants. Accumulation of this RNA peaks during embryonic and pupal periods. The osp cDNAs comprise two distinct classes based on alternative splicing patterns. The 5{prime} end of the longest cDNA was extended by PCR amplification. When hybridized to the osp walk, the 5{prime} extension verifies that Adh and Adh{sup r} are nested in osp and shows that osp has a transcription unit of {ge}74 kb. In situ hybridization shows that osp is expressed both maternally and zygotically. In the ovary, osp is transcribed in nurse cells and localized in the oocyte. In embryos, expression is most abundant in the developing visceral and somatic musculature. 55 refs., 11 figs., 1 tab.

Authors:
; ;  [1]
  1. Univ. of Cambridge (United Kingdom)
Publication Date:
OSTI Identifier:
543027
Resource Type:
Journal Article
Resource Relation:
Journal Name: Genetics; Journal Volume: 143; Journal Issue: 2; Other Information: PBD: Jun 1996
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; ALCOHOL DEHYDROGENASE; GENE AMPLIFICATION; STRUCTURE-ACTIVITY RELATIONSHIPS; GENE REGULATION; GENE MUTATIONS; SPLICING; TRANSCRIPTION; GENETIC MAPPING; CHROMOSOMES; CHROMOSOMAL ABERRATIONS; DROSOPHILA; ONTOGENESIS; MUTANTS; POLYMERASE CHAIN REACTION; DNA; EMBRYOS; IN-SITU HYBRIDIZATION; RNA; RECESSIVE MUTATIONS

Citation Formats

McNabb, S., Greig, S., and Davis, T. The alcohol dehydrogenase gene is nested in the outspread locus of Drosophila melanogaster. United States: N. p., 1996. Web.
McNabb, S., Greig, S., & Davis, T. The alcohol dehydrogenase gene is nested in the outspread locus of Drosophila melanogaster. United States.
McNabb, S., Greig, S., and Davis, T. 1996. "The alcohol dehydrogenase gene is nested in the outspread locus of Drosophila melanogaster". United States. doi:.
@article{osti_543027,
title = {The alcohol dehydrogenase gene is nested in the outspread locus of Drosophila melanogaster},
author = {McNabb, S. and Greig, S. and Davis, T.},
abstractNote = {This report describes the structure and expression of the outspread (osp) gene of Drosophila melanogaster. Previous work showed that chromosomal breakpoints associated with mutations of the osp locus map to both sides of the alcohol dehydrogenase gene (Adh), suggesting that Adh and the adjacent gene Adh{sup r} are nested in osp. We extended a chromosomal walk and mapped additional osp mutations to define the maximum molecular limit of osp as 119 kb. We identified a 6-kb transcript that hybridizes to osp region DNA and is altered or absent in osp mutants. Accumulation of this RNA peaks during embryonic and pupal periods. The osp cDNAs comprise two distinct classes based on alternative splicing patterns. The 5{prime} end of the longest cDNA was extended by PCR amplification. When hybridized to the osp walk, the 5{prime} extension verifies that Adh and Adh{sup r} are nested in osp and shows that osp has a transcription unit of {ge}74 kb. In situ hybridization shows that osp is expressed both maternally and zygotically. In the ovary, osp is transcribed in nurse cells and localized in the oocyte. In embryos, expression is most abundant in the developing visceral and somatic musculature. 55 refs., 11 figs., 1 tab.},
doi = {},
journal = {Genetics},
number = 2,
volume = 143,
place = {United States},
year = 1996,
month = 6
}
  • The gene that codes for Drosophila alcohol dehydrogenase was identified in a bacteriophage lambda library of genomic Drosophila DNA by using ADH cDNA cloned DNA as a probe. The DNA sequence of the protein encoding region was shown to be in agreement with the amino acid sequence of the ADH. Two intervening DNA sequences (introns) were identified within the protein encoding region. Both contained the 5' G-T and 3' A-G dinucleotides characteristic of intron boundaries of eukaryotic genes. On the basis of secondary structure predictions, the first 140 amino acid residues of Drosophila ADH are in an alternating ..beta..-sheet/..cap alpha..-helixmore » arrangement which is characteristic of the coenzyme binding domain of dehydrogenases. The smaller of the two introns interrupts the domain predicted to bind the adenine portion of the coenzyme.« less
  • The three forms of alcohol dehydrogenase (EC 1.1.1.1) within a given strain of Drosophila melanogaster are composed of similar, if not identical, peptide chains as shown by amino acid analysis and peptide fingerprinting. After feeding (carbonyl-/sup 14/C)nicotinamide to flies, label is associated with only two of the three forms in the ratio 1:2. Similarly, a fluorescent compound is associated with the same two forms. After purification of this compound and characterization of it by thin layer chromatography and mass spectroscopy, we conclude that the multiple forms of Drosophila alcohol dehydrogenase appear to be caused by the noncovalent binding of 1more » and 2 mol of an NAD-carbonyl compound addition complex to the enzyme.« less
  • The activity of alcohol dehydrogenase, 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 Adh{sup n2} 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 moremore » slowly than normal ethanol.l An examination of P element transformant lines with specific deletions in the 5{prime} regulatory DNA of the Adh gene showed that the DNA sequence between +604 and +634 of the start site of transcription from the distal promoter was essential for this induction. The DNA sequence between {minus}660 and about {minus}5,000 of the distal transcript start site was important for the down-regulation of the induction response.« less