Identification of DNA Sequences Required for the Regulation of Drosophila Alcohol Dehydrogenase Gene Expression

Posakony, James W.; Fischer, Janice A.; Maniatis, Tom

doi:10.1101/sqb.1985.050.01.063

Cited by 77 publications

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“…Various homoeodomain-binding sites were inserted just upstream by Jean-Paul Vincent (J.-P. Vincent and P.H.O'F., unpublished results). Responder 5 (pAF0) contains a proximal ADH promoter and structural gene 51 . Again, NP6 was inserted at the 5'-end of the promoter.…”

Section: Discussionmentioning

confidence: 99%

“…Homoeodomain-binding sites were inserted into unique restriction sites immediately upstream of the promoter. Responder 5 contains the Drosophila melanogaster proximal ADH promoter and structural gene starting at −386 nucleotides 51 . A PstI-EcoRI (blunted) fragment from the M13mp18 clone containing NP6 was inserted into the unique PstI site immediately upstream of the proximal ADH promoter.…”

Section: Discussionmentioning

confidence: 99%

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Activation and repression of transcription by homoeodomain-containing proteins that bind a common site

Jaynes

O’Farrell

1988

Nature

223

148

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The product of the fushi tarazu (ftz) gene is shown to be a site-dependent activator of transcription. In vitro-defined binding sites act as ftz-dependent enhancers in cultured cells. Another homoeodomain-containing protein, the engrailed gene product, competes for homoeodomainbinding sites and counteracts ftz activation.GENETIC analysis in Drosophila melanogaster has identified a group of regulatory genes that participate in embryonic pattern formation 1,2 . Many of these genes encode a 60 amino-acid sequence, the homoeodomain, that is highly conserved through evolution [3][4][5] . This domain contains a putative helix-turn-helix motif 6 such as that found in a number of bacterial regulators 7 , and possesses a sequence-specific DNA-binding activity 8,9 . Numerous regulatory interactions occur among homoeodomain-encoding genes during development. For example, a combination of segmentation and homoeotic gene products, many of which contain homoeodomains 1 , specify the developmental fates of cells in striped patterns 10,11 . Furthermore, homoeodomain-containing proteins (homoeodomain proteins) regulate both each other 12 and downstream (`cytodifferentiation' 13 ) genes. Consequently, it has been attractive to think of homoeodomain proteins as direct regulators of transcription. Due to the complexity of cross-regulatory interactions however, mutations altering or eliminating one homoeodomain protein generally change the expression patterns of many other regulators, confounding efforts to distinguish direct from indirect regulation. Additionally, suspected target genes contain large cis-acting control regions that respond, directly or indirectly, to complex combinations of regulators [14][15][16][17] . As a result, identification of target genes directly regulated by homoeodomain proteins has been problematic.Previous studies have documented instances in which homoeodomain proteins control transcription. The enhancer activity of a DNA fragment from the ftz gene, a homoeodomaincontaining member of the pair-rule class of segmentation genes, suggested that the ftz gene product (ftz) activates its own expression in the embryo 18 . In a different approach a Drosophila tissue culture system was used to show that the Ultrabithorax (Ubx) gene product induces expression of its own promoter, and inhibits the Antennapedia (Antp) (P1) promoter (M. Krasnow and D. S. Hogness, personal communication, see ref. 19 for summary). In these cases however, the complexity of the regulatory circuitry and of the responding control regions leaves open the possibility that the homoeodomain proteins produce these transcriptional effects through intermediaries. To circumvent the complexities of natural target genes, we chose to use binding sites identified in vitro to build homoeodomain-responsive promoters. We find that ftz is a potent activator of transcription from these promoters in cultured cells.During sequential subdivision of the Drosophila embryo to produce segmentally repeating patterns, combinations of regulatory gene prod...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Activation and repression of transcription by homoeodomain-containing proteins that bind a common site

Jaynes

O’Farrell

1988

Nature

223

148

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“…Should this be the case, it would almost certainly require restricting the activity of the region to the establishment of their common temporal activation during differentiation, as the two genes are regulated in quite different ways in the differentiated cells. Recent (49), and in fushi tarazu (22). In higher eucaryotes, the mouse cytochrome P1-450 gene has been found to have three functional domains, the most distal of which was between -1535 and -1265 bp.…”

Section: Methodsmentioning

confidence: 99%

Multiple regulatory elements in the intergenic region between the alpha-fetoprotein and albumin genes.

Godbout¹,

Ingram²,

Tilghman³

1986

Mol. Cell. Biol.

214

156

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Three enhancer elements spanning a distance of 7 kilobases have been found at the 5' end of the alpha-fetoprotein (AFP) gene. These elements were identified by transient expression assay after the introduction of a modified mouse AFP gene with variable amounts of 5' flanking sequence into a human hepatoma cell line, Hep G2. These regulatory elements function in a position-independent and orientation-independent manner that is typical of enhancers. All three elements will stimulate transcription from the promoter of the herpes simplex virus thymidine kinase gene. In Hep G2 cells, transcriptional activation from the heterologous promoter was approximately 25- to 50-fold higher than the basal levels obtained in the absence of AFP enhancer elements. In HeLa cells, the increase in thymidine kinase gene transcription varied from 6- to 14-fold, indicating that the enhancer elements exhibit some cell type specificity. Deletion analysis of the region proximal to the AFP transcription initiation site identified an essential region between 85 and 52 bases upstream of the site of initiation of transcription whose removal resulted in almost complete extinction of transcriptional activity. This region, which has been shown to be dispensable for transcription in HeLa cells, defines a second tissue-specific regulatory region in the gene.

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“…ADH enzyme activity per fly is 2-3 times higher in flies homozygous for AdhF alleles than in Adhs homozygotes and this is largely due to differences in ADH protein amounts (Gibson, 1972;Birley & Marson, 1991;Maroni et a!., 1982) but the higher catalytic efficiency of ADH-F (Winberg et a!., 1985) also has an effect. ADH activity and tissue distribution are directly affected by genetic variants at the structural gene locus, and are regulated by cis and trans acting elements (Goldberg et a!., 1983;Posakony et a!., 1985;Fischer & Maniatis, 1986;Corbin & Maniatis, 1990). Clarke & Whitehead (1984) have argued that the structural and regulatory contributions to ADH variation are likely to have co-evolved as a linked group of selectively interacting factors.…”

Section: Introductionmentioning

confidence: 99%

The alcohol dehydrogenase polymorphism in natural populations of Drosophila melanogaster: ADH activity variation restriction site polymorphism and the Adh cline

Jiang

Gibson

1992

Heredity

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Alcohol dehydrogenase activity has been measured in 186 iso-second chromosome lines -104 from seven Australian populations and 82 from six Chinese populations. Restriction endonuclease variation in the Adh gene region in these lines has previously been described (Jiang & Gibson, 1991). The mean ADH activity of AdhF and Adhs lines was significantly higher in the Chinese samples than in the Australian samples. In each population on both continents the mean activity of the AdhF lines is significantly higher than that of the Adhs lines. Six lines homozygous for a thermostability variant, AdhF (detected in four of the Chinese populations), had intermediate levels of ADH activity and protein amount. In a subset of the lines with the highest and lowest levels of ADH, there was a correlation of 0.69 between ADH activity and ADH CRM. None of the restriction site variants was consistently associated with the amount of ADH activity. Associations between BamHI (-7.2), the Adh polymorphism and ADH activity suggest that there are modifiers of ADH 5' to the gene. The deletion (0.2) at position -2.8 on the restriction map (Jiang & Gibson, 1991) was associated with increased levels of ADH, activity in Adhs lines from China. Two unique insertions in the gene region were associated with low activity in AdhF lines and a null activity allele had a deletion removing most of exon 2. A single line with a duplication of a part of the Adh coding region and of the 5' regulatory section had relatively high ADH activity. Considering all the data, the main factor affecting ADH activity levels in populations is the frequency of AdhF.

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Identification of DNA Sequences Required for the Regulation of Drosophila Alcohol Dehydrogenase Gene Expression

Cited by 77 publications

References 0 publications

Activation and repression of transcription by homoeodomain-containing proteins that bind a common site

Activation and repression of transcription by homoeodomain-containing proteins that bind a common site

Multiple regulatory elements in the intergenic region between the alpha-fetoprotein and albumin genes.

The alcohol dehydrogenase polymorphism in natural populations of Drosophila melanogaster: ADH activity variation restriction site polymorphism and the Adh cline

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