scholarly journals Molecular characterization of the Drosophila melanogaster urate oxidase gene, an ecdysone-repressible gene expressed only in the malpighian tubules.

1990 ◽  
Vol 10 (10) ◽  
pp. 5114-5127 ◽  
Author(s):  
L L Wallrath ◽  
J B Burnett ◽  
T B Friedman
Keyword(s):  
Drosophila Melanogaster ◽  
Amino Acid ◽  
Regulatory Element ◽  
Malpighian Tubule ◽  
Amino Acid Sequences ◽  
Malpighian Tubules ◽  
P Element ◽  
Urate Oxidase ◽  
Base Pairs ◽  
Oxidase Gene

The urate oxidase (UO) gene of Drosophila melanogaster is expressed during the third-instar larval and adult stages, exclusively within a subset of cells of the Malpighian tubules. The UO gene contains a 69-base-pair intron and encodes mature mRNAs of 1,224, 1,227, and 1,244 nucleotides, depending on the site of 3' endonucleolytic cleavage prior to polyadenylation. A direct repeat, 5'-AAGTGAGAGTGAT-3', is the proposed cis-regulatory element involved in 20-hydroxyecdysone repression of the UO gene. The deduced amino acid sequences of UO of D. melanogaster, rat, mouse, and pig and uricase II of soybean show 32 to 38% identity, with 22% of amino acid residues identical in all species. With use of P-element-mediated germ line transformation, 826 base pairs 5' and approximately 1,200 base pairs 3' of the D. melanogaster UO transcribed region contain all of the cis elements allowing for appropriate temporal regulation and Malpighian tubule-specific expression of the UO gene.

Molecular characterization of the Drosophila melanogaster urate oxidase gene, an ecdysone-repressible gene expressed only in the malpighian tubules

1990 ◽  
Vol 10 (10) ◽  
pp. 5114-5127
Author(s):  
L L Wallrath ◽  
J B Burnett ◽  
T B Friedman
Keyword(s):  
Drosophila Melanogaster ◽  
Amino Acid ◽  
Regulatory Element ◽  
Malpighian Tubule ◽  
Amino Acid Sequences ◽  
Malpighian Tubules ◽  
P Element ◽  
Urate Oxidase ◽  
Base Pairs ◽  
Oxidase Gene

The urate oxidase (UO) gene of Drosophila melanogaster is expressed during the third-instar larval and adult stages, exclusively within a subset of cells of the Malpighian tubules. The UO gene contains a 69-base-pair intron and encodes mature mRNAs of 1,224, 1,227, and 1,244 nucleotides, depending on the site of 3' endonucleolytic cleavage prior to polyadenylation. A direct repeat, 5'-AAGTGAGAGTGAT-3', is the proposed cis-regulatory element involved in 20-hydroxyecdysone repression of the UO gene. The deduced amino acid sequences of UO of D. melanogaster, rat, mouse, and pig and uricase II of soybean show 32 to 38% identity, with 22% of amino acid residues identical in all species. With use of P-element-mediated germ line transformation, 826 base pairs 5' and approximately 1,200 base pairs 3' of the D. melanogaster UO transcribed region contain all of the cis elements allowing for appropriate temporal regulation and Malpighian tubule-specific expression of the UO gene.

scholarly journals Isolation and characterization of the Beadex locus of Drosophila melanogaster: a putative cis-acting negative regulatory element for the heldup-a gene.

1984 ◽  
Vol 4 (7) ◽  
pp. 1343-1353 ◽  
Author(s):  
W W Mattox ◽  
N Davidson
Keyword(s):  
Drosophila Melanogaster ◽  
Base Pair ◽  
Regulatory Element ◽  
P Element ◽  
Functional Domain ◽  
Lambda Phage ◽  
Base Pairs ◽  
Negative Regulatory Element ◽  
Cis Acting ◽  
Isolation And Characterization

We isolated recombinant lambda phage clones spanning 49 kilobases of DNA which contain the Beadex and heldup-a loci of Drosophila melanogaster. These cloned DNAs were used to analyze the structure of eight dominant mutant alleles of the Beadex locus which show increased gene activity. A region, only 700 base pairs in length, is altered in each of these mutants. Six of the mutations have DNA insertions within this segment. Most of these insertions resemble retrovirus-like transposable elements. In one case (Beadex2) the inserted sequences are homologous to the gypsy transposon family. The other two Beadex alleles were induced by hybrid dysgenesis and suffered deletions which included at least part of the 700-base-pair segment. These deletions appear to have resulted from imprecise excision or deletion of a nearby P element found in the wild-type parental strain. Analysis of one heldup-a allele (heldup-aD30r) indicates that a similar P element-mediated event is responsible for this lesion. In this mutant, deletion of sequences no more than 1,600 base pairs from the Beadex locus accompanies the loss of heldup-a function. The deleted sequences in heldup-aD30r include the entire 700-base-pair segment within which at least part of the Beadex locus resides, yet these flies have no Beadex phenotype. This indicates that a functional heldup-a gene is necessary for expression of the Beadex phenotype. Together, these results suggest that the Beadex functional domain is contained within a short segment of DNA near the heldup-a gene and support the hypothesis that the Beadex locus functions as a cis-acting negative regulatory element for the heldup-a gene.

scholarly journals Isolation and characterization of the Beadex locus of Drosophila melanogaster: a putative cis-acting negative regulatory element for the heldup-a gene

1984 ◽  
Vol 4 (7) ◽  
pp. 1343-1353
Author(s):  
W W Mattox ◽  
N Davidson
Keyword(s):  
Drosophila Melanogaster ◽  
Base Pair ◽  
Regulatory Element ◽  
P Element ◽  
Functional Domain ◽  
Lambda Phage ◽  
Base Pairs ◽  
Negative Regulatory Element ◽  
Cis Acting ◽  
Isolation And Characterization

We isolated recombinant lambda phage clones spanning 49 kilobases of DNA which contain the Beadex and heldup-a loci of Drosophila melanogaster. These cloned DNAs were used to analyze the structure of eight dominant mutant alleles of the Beadex locus which show increased gene activity. A region, only 700 base pairs in length, is altered in each of these mutants. Six of the mutations have DNA insertions within this segment. Most of these insertions resemble retrovirus-like transposable elements. In one case (Beadex2) the inserted sequences are homologous to the gypsy transposon family. The other two Beadex alleles were induced by hybrid dysgenesis and suffered deletions which included at least part of the 700-base-pair segment. These deletions appear to have resulted from imprecise excision or deletion of a nearby P element found in the wild-type parental strain. Analysis of one heldup-a allele (heldup-aD30r) indicates that a similar P element-mediated event is responsible for this lesion. In this mutant, deletion of sequences no more than 1,600 base pairs from the Beadex locus accompanies the loss of heldup-a function. The deleted sequences in heldup-aD30r include the entire 700-base-pair segment within which at least part of the Beadex locus resides, yet these flies have no Beadex phenotype. This indicates that a functional heldup-a gene is necessary for expression of the Beadex phenotype. Together, these results suggest that the Beadex functional domain is contained within a short segment of DNA near the heldup-a gene and support the hypothesis that the Beadex locus functions as a cis-acting negative regulatory element for the heldup-a gene.

Amino-Acid Metabolism in Locust Tissues

1957 ◽  
Vol 34 (2) ◽  
pp. 276-289
Author(s):  
B. A. KILBY ◽  
ELISABETH NEVILLE
Keyword(s):  
Amino Acid ◽  
Amino Acid Metabolism ◽  
Acid Metabolism ◽  
Schistocerca Gregaria ◽  
Fat Body ◽  
Malpighian Tubule ◽  
Malpighian Tubules ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Mitochondrial Fractions

1. Homogenates of fat-body of Schistocerca gregaria Forsk. were shown to catalyse transamination reactions between α-ketoglutarate and numerous α-amino acids. The aspartate/glutamate and alanine/glutamate transaminases were the most active. They were present in both the ‘soluble’ and the mitochondrial fractions of fat-body cells and also in Malpighian tubules and mid-gut wall. The other transaminases in the fat-body were confined to the mitochondrial fraction. 2. Fat-body, Malpighian tubule and mid-gut wall homogenates were able to convert glutamic acid into glutamine, a compound which could also act as an amino-group donor in some transamination reactions. 3. A glutamate-cytochrome c reductase system which involved diphosphopyridine nucleotide was present in fat-body. 4. Fat-body contained an active arginase, but urease could not be detected. A D-amino-acid oxidase was present, together with a less active L-amino-acid oxidase. 5. In general, it appears that amino-acid metabolism in the locust resembles that in higher animals.

scholarly journals Sequences involved in temperature and ecdysterone-induced transcription are located in separate regions of a Drosophila melanogaster heat shock gene.

1986 ◽  
Vol 6 (2) ◽  
pp. 663-673 ◽  
Author(s):  
E Hoffman ◽  
V Corces
Keyword(s):  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Dna Sequences ◽  
Germ Line ◽  
Initiation Site ◽  
P Element ◽  
Heat Shock Gene ◽  
Base Pairs ◽  
Consensus Sequences ◽  
Shock Gene

The transcriptional regulation of the Drosophila melanogaster hsp27 (also called hsp28) gene was studied by introducing altered genes into the germ line by P element-mediated transformation. DNA sequences upstream of the gene were defined with respect to their effect on steroid hormone-induced and heat-induced transcription. These two types of control were found to be separable; the sequences responsible for 80% of heat-induced expression were located more than 1.1 kilobases upstream of the RNA initiation site, while the sequences responsible for the majority of ecdysterone induction were positioned downstream of the site at -227 base pairs. We have determined the DNA sequence of the intergenic region separating hsp23 and hsp27 and have located putative heat shock and ecdysterone consensus sequences. Our results indicate that the heat shock promoter of the hsp27 gene is organized quite differently from that of hsp70.

scholarly journals Genes for low-molecular-weight heat shock proteins of soybeans: sequence analysis of a multigene family.

1985 ◽  
Vol 5 (12) ◽  
pp. 3417-3428 ◽  
Author(s):  
R T Nagao ◽  
E Czarnecka ◽  
W B Gurley ◽  
F Schöffl ◽  
J L Key
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Heat Shock ◽  
Dna Sequences ◽  
Regulatory Element ◽  
Amino Acid Sequences ◽  
Striking Similarity ◽  
Low Molecular Weight ◽  
Genes Encoding ◽  
Flanking Regions

Soybeans, Glycine max, synthesize a family of low-molecular-weight heat shock (HS) proteins in response to HS. The DNA sequences of two genes encoding 17.5- and 17.6-kilodalton HS proteins were determined. Nuclease S1 mapping of the corresponding mRNA indicated multiple start termini at the 5' end and multiple stop termini at the 3' end. These two genes were compared with two other soybean HS genes of similar size. A comparison among the 5' flanking regions encompassing the presumptive HS promoter of the soybean HS-protein genes demonstrated this region to be extremely homologous. Analysis of the DNA sequences in the 5' flanking regions of the soybean genes with the corresponding regions of Drosophila melanogaster HS-protein genes revealed striking similarity between plants and animals in the presumptive promoter structure of thermoinducible genes. Sequences related to the Drosophila HS consensus regulatory element were found 57 to 62 base pairs 5' to the start of transcription in addition to secondary HS consensus elements located further upstream. Comparative analysis of the deduced amino acid sequences of four soybean HS proteins illustrated that these proteins were greater than 90% homologous. Comparison of the amino acid sequence for soybean HS proteins with other organisms showed much lower homology (less than 20%). Hydropathy profiles for Drosophila, Xenopus, Caenorhabditis elegans, and G. max HS proteins showed a similarity of major hydrophilic and hydrophobic regions, which suggests conservation of functional domains for these proteins among widely dispersed organisms.

scholarly journals P-element-induced control mutations at the r gene of Drosophila melanogaster.

1985 ◽  
Vol 5 (10) ◽  
pp. 2567-2574 ◽  
Author(s):  
S Tsubota ◽  
M Ashburner ◽  
P Schedl
Keyword(s):  
Drosophila Melanogaster ◽  
The Other ◽  
P Element ◽  
Female Sterility ◽  
R Gene ◽  
Temporal Expression ◽  
Base Pairs ◽  
P Elements ◽  
Adult Females ◽  
Regulatory Mutations

The P-M hybrid dysgenesis system was used to produce five putative regulatory mutations at the rudimentary locus, r. All five mutations were the result of insertions at the 5' end of the gene, upstream of the proposed start of transcription. All of the mutants displayed a leaky wing phenotype, and four of the mutants showed an uncoupling of the wing and female-sterility phenotypes, suggesting that they altered the normal spatial and temporal expression of the r gene. Four of the insertions were P elements. The fifth insertion, which was larger than an intact P element, consisted of a small P element connected to non-P-element DNA. Two of the mutants produced very little r transcript in adult females and were clustered 80 to 150 base pairs upstream of the start of transcription. The other three mutants had higher levels of r transcript in adult females and were clustered 440 to 500 base pairs upstream of the start of transcription. All of the data suggest that the insertions are in a 5' noncoding region of the r gene involved in the control of its spatial and temporal expression.

scholarly journals Homology Requirements for Targeting Heterologous Sequences During P-Induced Gap Repair in Drosophila melanogaster  1

Genetics ◽  
1997 ◽  
Vol 147 (2) ◽  
pp. 689-699 ◽  
Author(s):  
Tammy Dray ◽  
Gregory B Gloor
Keyword(s):  
Drosophila Melanogaster ◽  
Point Mutations ◽  
Double Strand Breaks ◽  
Yellow Gene ◽  
P Element ◽  
White Gene ◽  
Base Pairs ◽  
Strand Breaks ◽  
Distal Point ◽  
White Locus

The effect of homology on gene targeting was studied in the context of P-element-induced double-strand breaks at the white locus of Drosophila melanogaster. Double-strand breaks were made by excision of P-whd, a P-element insertion in the white gene. A nested set of repair templates was generated that contained the 8 kilobase (kb) yellow gene embedded within varying amounts of white gene sequence. Repair with unlimited homology was also analyzed. Flies were scored phenotypically for conversion of the yellow gene to the white locus. Targeting of the yellow gene was abolished when all of the 3′ homology was removed. Increases in template homology up to 51 base pairs (bp) did not significantly promote targeting. Maximum conversion was observed with a construct containing 493 bp of homology, without a significant increase in frequency when homology extended to the tips of the chromosome. These results demonstrate that the homology requirements for targeting a large heterologous insertion are quite different than those for a point mutation. Furthermore, heterologous insertions strongly affect the homology requirements for the conversion of distal point mutations. Several aberrant conversion tracts, which arose from templates that contained reduced homology, also were examined and characterized.

Structure and expression of ubiquitin gene transcripts in pine

1995 ◽  
Vol 25 (1) ◽  
pp. 1-7 ◽  
Author(s):  
M. Carol Alosi Carter ◽  
Rima M. Kulikauskas ◽  
Roderic B. Park
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Northern Blot Analysis ◽  
Amino Acid Sequences ◽  
Base Pairs ◽  
Phloem Tissue ◽  
Computer Aided Analysis ◽  
Computer Aided ◽  
Gene Transcripts ◽  
A Minor

Clones encoding polyubiquitin proteins were isolated from a cDNA library derived from Pinussabiniana phloem tissue. Two different polyubiquitin clones were sequenced. The amino acid sequences of the clones were compared with angiosperm polyubiquitin sequences and with corresponding sequences reported for animal, fungal, and protist polyubiquitins. A computer-aided analysis showed (i) that pine and angiosperm polyubiquitin amino acid sequences correspond perfectly, (ii) that plant polyubiquitins differ from animal polyubiquitins by three amino acids, and (iii) that fungal and protist polyubiquitins are variable, differing by one to eight amino acids from higher organisms. The expression of ubiquitin was studied in bark, cone, needle, phloem, root, and xylem tissues of pine by Northern blot analysis. In all of these tissues, transcripts of about 1000 base pairs were observed. A minor species of 1200 base pairs was seen in longer exposures of autoradiograms. Ubiquitin transcripts were more abundant (in relationship to total RNA) in phloem, cones, and roots than in bark, needles, or xylem.

scholarly journals The molecular correlates of organ loss: the case of insect Malpighian tubules

2015 ◽  
Vol 11 (5) ◽  
pp. 20150154 ◽  
Author(s):  
Xiangfeng Jing ◽  
Thomas A. White ◽  
Xiaowei Yang ◽  
Angela E. Douglas
Keyword(s):  
Amino Acid ◽  
Amino Acid Metabolism ◽  
Essential Role ◽  
Acyrthosiphon Pisum ◽  
Acid Metabolism ◽  
Malpighian Tubule ◽  
Malpighian Tubules ◽  
Pea Aphid ◽  
Whole Genome ◽  
Central Importance

Malpighian tubules play an essential role in excretion, osmoregulation and immunity of most insects. Exceptionally, aphids lack Malpighian tubules, providing the opportunity to investigate the fate of genes expressed in an organ that has undergone evolutionary reduction and loss. Making use of the sequenced genomes of Drosophila melanogaster and the pea aphid Acyrthosiphon pisum , we demonstrated that more than 50% of Drosophila genes expressed specifically in the Malpighian tubules had orthologues in the pea aphid genome and that most of the pea aphid orthologues with detectable expression were identified in the gut transcriptome. Relative to the whole genome, genes functioning in amino acid metabolism are significantly over-represented among the pea aphid orthologues of Malpighian tubule genes, likely reflecting the central importance of amino acid acquisition and metabolism in aphids. This study demonstrates that the evolutionary loss of a key insect organ, the Malpighian tubules, has not been associated with the coupled loss of molecular functions.

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