BG:DS01486.8 , DmADH
Dicistronic transcript isoform(s) appear to be relatively rare based on RNA-Seq and/or EST data.
Gene model reviewed during 5.52
Low-frequency RNA-Seq exon junction(s) not annotated.
Gene model reviewed during 5.55
In vitro transcribed protein from a genomic Adh clone (gAC1 and sAC1) which was bound by an immobilized Adh antibody was run on a gel to show one protein product with an approximate size of 24kD. No Adh protein was recovered from an Actin clone or without the addition of DNA.
Click to get a list of regulatory features (enhancers, TFBS, etc.) and gene disruptions (point mutations, indels, etc.) within or overlapping Dmel\Adh using the Feature Mapper tool.
As the concentration of ethanol in the diet is increased, the amount of Adh protein in the midgut increases. None is observed in the foregut or hindgut.
Monoclonal antibodies MMBB8 and LLBE8 were used to analyse the temporal and tissue-specific patterns of Adh gene expression. In the early stages of oogenesis, small amounts of Adh protein are detectable in the cystocytes. At the beginning of vitellogenesis, Adh protein is located mainly in the nurse cells. During late oogenesis, multiple Adh protein-positive bodies of varying size appear in the ooplasm. Adh protein is compartmentalized within the yolk or β-spheres.
In tissues such as fat body, gastric caeca, and adult cardiac valve the patterns of Adh RNA and protein expression are identical. Other tissues such as oocytes, nurse cells, imaginal discs, and brain have the same or lower levels of RNA but little or no Adh protein.
GBrowse - Visual display of RNA-Seq signalsView Dmel\Adh in GBrowse 2
Please Note FlyBase no longer curates genomic clone accessions so this list may not be complete
Please Note This section lists cDNAs and ESTs that fall within the genomic extent of the gene model, which may include cDNAs and ESTs of genes within introns, or of overlapping genes. Please see GBrowse for alignment of the cDNAs and ESTs to the gene model.
For each fully sequenced cDNA the DGRC maintains various forms of the cDNA (e.g tagged or untagged) in several different host vectors for subsequent cloning and expression in Drosophila and Drosophila cell lines.
Source for identity of: Adh CG3481
Dicistronic annotation CG32954 split out into separate annotations for each open reading frame, CG3481 and CG3484, in release 4.2 of the genome annotation. CG3481 corresponds to Adh and CG3484 corresponds to Adhr.
Full-length Adh transgenes are silenced post-transcriptionally at high copy number or by a pulsed increase over a threshold. Production of 21-25 bp length sense and antisense RNAs homologous to Adh is correlated with this process.
A non-transcribed segment in the Adh regulatory region was found to be one of the sequences required for homology recognition in the phenomenon of cosuppression.
Transgene coplacement studies indicate that the correlation in specific activity between D.melanogaster and D.affinidisjuncta Adh genes that occupy the same position is high in both larvae and adults.
5' end RACE of the Adhr transcript from adults indicates that Adhr is transcribed as a dicistronic mRNA from the Adh distal promoter. RACE experiment with total RNA from embryos showed the embryonic Adhr transcript is also dicistronic and is transcribed from the proximal Adh promoter. Mutations that affect Adh transcripts also affect Adhr transcripts. Sedimentation profiles of polysomes and RNA analysis indicates the Adhr open reading frame of the dicistronic transcript is being translated and strongly suggests that Adhr translation is initiated by internal initiation in the intergenic region between Adh and Adhr coding regions. Adhr protein can be detected and is shown to co-localise with Adh. The level of Adh and Adhr transcripts in su(f) mutant and wild type background demonstrates the accumulation of the dicistronic messenger is controlled by a temperature-sensitive post-transcriptional mechanism.
Molecular replacement and data from crystallographically refined 3D determination structures confirm the position of Ser 139. Results suggest Ser 139 is directly involved in the catalytic reaction.
NS1 region is located approximately 300bp upstream of the larval cap site and acts like an enhancer. Adh protein is not expressed when the sequence is deleted, expression is restored by placing a second Adh gene with an intact enhancer elsewhere on the same plasmid. The interactions between the enhancer and expression from a proximal and distal promoter are investigated.
The excision reactions of P1\cre/P1\loxP and the related Scer\FLP1/Scer\FRT system are used to create lines in which transgenes, Adh and Daff\Adh, are at exactly allelic sites in homologous chromosomes.
A 3D model of Adh using the tertiary structures has been generated, using molecular information about mutant alleles, to provide additional information about Adh catalysis and the stability of Adh dimers.
mRNA levels do not increase at adult day 5 in strain showing extended longevity phenotype (ELP).
The tandem Adh promoters are differentially transcribed in the embryo owing to critical differences in the core promoter elements. Reconstituted differential Adh promoter transcription in vitro provides evidence that selective Adh promoter utilization is mediated by a specific Tbp-TAF complex in combination with TfIIA. TAFs in the Tbp complex are required for discrimination between the Adh distal and proximal initiator elements.
Wild type larvae are most tolerant to ethanol, then methanol, n-propanol and n-butanol in descending order. For Adh-deficient larvae the toxicity of methanol was least, then ethanol, n-propanol and n-butanol in that order. Toxicity for this stage of the larval period is related to chain length of the alcohol, if the alcohol could not be degraded by the Adh system.
Bending of DNA can inhibit transcription by affecting the binding of at least one of the general transcription factors. A tight bend present in a 245bp minicircular DNA reduces transcription from the Adh distal promoter. Repression occurs, at least in part, because the general factor TFIID is unable to bind to this bent DNA.
Isolated as a rhythmically expressed transcript in fly heads. Transcript shows a single daily peak and a single daily trough in expression, highest expression is in the 'morning'. Adh expression oscillation is dependent on a light-dark cycle, on timed feeding and on the function of the per gene.
The homologous genomic region containing Adh and Adhr is analysed. Ka and Ks values are determined (Ks values for Adh are significantly lower than values for Adhr) and amino acid comparisons reveal conserved regions shared by Adh and Adhr which have been assigned to known functional domains.
Adh activity in 71 Drosophila species is assayed to determine if the protein plays a key role in the adaptation of species to substrates undergoing alcoholic fermentation.
The in situ localization of Adh transcripts in different species reveals evolved regulatory differences in spatially restricted expression.
The srp gene encodes a transcription factor that binds a conserved sequence element of the larval promoter of the Adh gene. The srp-binding sites of the D.melanogaster and D.mulleri Adh larval promoters function as positive control elements.
The function of residues Tyr152 and Lys156 has been studied and the enzymatic properties of the mutants determined.
Mutation of the Adh gene suggests that Tyr152 and Lys156 are involved in catalysis and Gly130, Gly133 and Gly184 contribute substantially to the structure of the active form.
Adh protein either copurifies with a subtilisin type protease or may have protease activity per se.
The effect of dietary ethanol on the ultrastructure of wild type and Adh mutant larvae was studied. The midgut and hindgut showed ethanol-induced subcellular damage, disrupting mitochondria and endoplasmic reticulum, reducing glycogen rosettes and protein granules, increasing autophagic vacuoles and causing unusual myelin whorls, with Adh mutant having greater sensitivity to ethanol.
Accessible chromatin structures are found in the Adh proximal promoter, distal promoter and adult enhancer region just prior to and during Adh transcription in the fat body. A nucleosome positioning element between the distal promoter and the Adh adult enhancer can function via DNA-core histone interactions alone.
Aldehyde dehydrogenase activities from Adh and Aldh gene products are selectively inhibited by cyanimide or acetone, respectively. Although larvae and adults use different aldehyde dehydrogenase activities to detoxify acetaldehyde (from Adh and Aldh encoded enzymes, respectively) both activities are cytosolic.
Results are opposed to those of Lietaert, Experientia 38:651 and Lietaert, Experientia 41:57 , who concluded that the aldehyde dehydrogenase activity was mainly in the mitochondria.
Phylogenetic relationships in Drosophila are studied using the Alcohol dehydrogenase locus in several species.
The tyrosine at the invariant amino acid position 152 is essential for the activity of the Adh enzyme.
To the contrary of what occurs in larvae, the short-term modulation of the enzyme activities involved in the metabolism of alcohols does not appear to be a major mechanism used by adults to respond to the presence of ethanol, 2-propanol and their respective in vivo oxidised derivatives in the medium. Adults ability to move to non-toxic feeding sites seems to be successful enough to avoid the toxic effects of alcohols.
A sequence specific DNA binding factor, Trl, has binding sites that flank the distal promoter elements of Adh involved in transcription initiation. Trl acts as a repressor of expression acting at transcription initiation and binding requires an intact 10bp motif.
flies lacking Adh protein rapidly become intoxicated and eventually die on exposure to ethanol.
Confers resistance to ethanol.
Four alleles, AdhF, AdhS, Adhn4 and Adh71k were tested for oviposition site preference and first instar larval food preference in multiple choice tests between different media. Strains showed significantly different patterns.
A negative regulatory element in the AAE binds the adult enhancer factor 1 (Aef1). The Aef1 binding site, Dmul\Adh1 AAE and Yp1 gene fat body enhancer are related to a sequence recognised by the mammalian transcription factor C/EBP and a liver specific regulatory element of the human Adh gene. DNase I footprinting experiments reveal that Aef1 and C/EBP compete for adjacent binding sites in the fat body enhancers, Aef1 can displace bound C/EBP from its sites.
Adh null activity alleles extracted from a number of natural populations in Tasmania are molecularly similar.
High resolution analysis of chromatin structure and helix distortion around regions required for distal transcription of Adh revealed apparent coordinate assembly of a large cooperative complex of proteins interacting with the distal promoter, the positioned nucleosome, the enhancer of the distal promoter and each other.
A wild type 3.2 kb Adh gene fragment has been inserted into an Adh- strain in multiple chromosomal locations by P-element mediated transformation, identifying tissue specific position effects that possibly reflect differing chromatin organization.
Limited regions of Adh are especially sensitive to proteolysis, results suggest the possibility of an association between the enzyme active site and the sensitive site(s).
The reducing activity of the Adh enzyme, which transforms acetaldehyde into ethanol, plays an essential role in the detoxification of acetaldehyde.
Analysis of a number of different isogenic lines containing the natural polymorphism 'upside down triangle 2' suggests an association with the polymorphism and higher levels of Adh protein.
Comparison of CpG distribution in the coding region of 121 genes from six species supports the mCpG mutational hotspot explanation of CpG suppression in methylated species at position II-III and III-I.
Adaptation to ethanol may not be strictly a function of the level of Adh pprotein activity.
Two enhancer sequences, AAE and ALE, are required for the efficient expression of the distal and proximal promoters, respectively. The ALE segment can be sub- divided into three segments which act synergistically. ALE is ineffective in adults because transcription takes place from the distal promoter: transcriptional interference. The interactions of transcriptional interference at the molecular level are unknown. The ALE is involved in the down regulation of the ethanol induction response, the AAE is unresponsive to ethanol.
Previous site directed in vitro mutagenesis experiments have demonstrated the average difference in Adh protein level between the fast and slow allozymic classes may or may not be due to linkage disequilibrium between the amino acid replacement site and the polymorphism present in the insert fragment, a silent substitution at nucleotide 1443.
Larval expression is dependent on a 53bp sequence located upstream of the larval transcription start site.
Transcribed larval mRNA was measured from the autosomal Adh gene and the X chromosomal Dpse\Hsp83 gene, both carried on the same P-element construct. The compensation behaviour of both of the transposed genes was determined by their new chromosomal environment.
A 15 bp positive cis-acting element nearly 500 bp upstream of the distal Adh RNA start site and a 61 bp negative cis-acting element upstream and adjacent to the enhancer behave as promoter elements in the tissue culture system.
Flies trisomic for a quarter of the length of 2L have diploid Adh enzyme activity and mRNA levels. By subdividing the trisomic chromosome a region exerting an inverse regulatory effect and a region exerting a direct gene dosage response on Adh was found. When present simultaneously the regions cancel each other out to yield diploid levels of Adh enzyme activity and mRNA.
Transcription from the Adh proximal promoter is regulated by a far upstream enhancer and at least two elements near the transcription start site. The enhancer is tissue specific, and includes at least two discrete regions. Each of the identified regulatory elements is sufficient for low levels of Adh gene expression in larval tissues, but maximal expression requires the entire set.
Chromatin at the Adh distal promoter displays an ordered but different conformation in different cell types. In Adh- cells sequences between -40 to +30 of the distal RNA initiation site exist as a DNA linker between nucleosomes. In Adh+ cells a longer linker DNA, -140 to +30, is bound in a multi-protein transcription initiation complex. These mutually exclusive patterns of DNA- protein interactions suggest a model for organizing alternative chromatin structure associated with gene regulation.
Analysis of ENU-induced Adh mutations demonstrates that ENU produces primarily GC to AT transitions, also transversions and multilocus deficiencies.
The regions between +604bp and +634bp, and between -600bp and -5000bp significantly affect the induction of Adh by ethanol.
The tissue specific expression of the D.orena Adh gene in D.melanogaster is similar to D.melanogaster but at lower levels. The Adh genes share conserved DNAse protected sequences with respect to position and sequence. Upstream regions show a mosaic of similar and dissimilar sequences.
Northern blot analysis and gel electrophoresis of transformant D.melanogaster larvae carrying the D.affinidisjuncta and D.grimshawi Adh gene show comparable high levels of expression and broader tissue distribution of Adh expression, transformants carrying the D.hawaiiensis Adh gene show reduced levels of each.
Results suggest the switch from Adh proximal to Adh distal promoter is regulated by the stage-specific activation of the distal promoter and the subsequent repression of the proximal promoter by transcriptional interference.
Three Adh antibodies were characterized and immunoblotting assays found that they cross-react to Adh genes from D.melanogaster, D.bocqueti, D.erecta, D.teissieri and D.lebanonensis. Adh specific activity in different larval organs was found to be similar whereas protein distribution varies substantially.
Studies of the flux of ethanol into lipid suggested that more than 75% of the oxidation of acetaldehyde in wild type larvae is catalysed by the Adh product, the remaining ethanol is oxidized by the Aldh product.
Gene conversion and unequal crossover have been studied in flies carrying a construct that contains tandem copies of Adh- genes. Southern blots of Adh+ variants suggests that five were generated by gene conversion as the size of the P-element insert is unchanged and one was generated by unequal exchange as the size of the Adh cluster has been changed.
Maternally inherited Adh transcripts decay rapidly. Zygotic expression of Adh RNA begins after germ band retraction. Distal and proximal promoters drive expression in the fat body. At 15 hours proximal promoter expression is seen in the gut, at this time distal expression in the fat body has ceased. Proximal transcript levels decline at the end of larval development, accompanied by a transient accumulation of distal transcripts predominantly in the fat body.
Analysis of three populations by restiction mapping in Adh provides evidence of Founder effects in the most Northern populations. There are signs of population differentiation among the samples, but the similarities between the populations indicates extensive migration. This suggests natural selection plays a role in maintaining the cline.
Daff\Adh is expressed at comparable levels in D.melanogaster and D.affinidisjuncta, and tissue and stage specificity of expression is similar in the two species. In some details expression of Daff\Adh in D.melanogaster resembles that of Daff\Adh in D.affinidisjuncta.
Daff\Adh and Dhaw\Adh display markedly different levels of alcohol dehydrogenase in the larval midgut and Malpighian tubules. Comparison of the expression of Daff\Adh and Dhaw\Adh in transgenes in D.melanogaster demonstrates that the tissue specific levels of alcohol dehydrogenase are characteristic of the genes themselves. Demonstrable differences in cis-dominant regulatory information are sufficient to account for the observed regulatory variation.
Not expressed in SL2 tissue culture cells, but transfected cloned gene is.
Specific activity of the Adh product changes with development, with peaks at the end of the third larval instar and about four days after eclosion.
The Adh product is not a metalloenzyme but, paradoxically, is inhibited by certain metal ion chelators, e.g. pyrazole.
Adh may play a metabolic role independent of alcohol detoxification, i.e. in the metabolism of higher alcohols.
Specific activity of the Adh product changes with development, with peaks at the end of the third larval instar and about four days after eclosion. Half life of AdhF product in vivo estimated as 55.3 hours.
Ethanol tolerance usually correlated with Adh product activity.
Adh protein is maternally inherited by embryos and resulting larvae.
flies lacking Adh protein rapidly become intoxicated and eventually die on exposure to ethanol. However, ethanol sensitivity is complex since even Adh nulls are more resistant to ethanol when young than when old.
Adh+ flies are killed by low concentrations of unsaturated secondary alcohols (e.g. 1-penten-3-ol; 1-pentyn-3-ol) but not by unsaturated primary alcohols (e.g. 1-penten-1-ol), presumably due to the formation of toxic ketones. This allows the chemical selection of Adh nulls.