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V G Corces

Publications and source records attributed to V G Corces.

71 records · Page 4Linked to original sources
PubMed1988

Expression of an activated ras gene causes developmental abnormalities in transgenic Drosophila melanogaster.

We describe the developmental effects of the expression of a normal and mutated Drosophila ras gene ras2 in transgenic Drosophila. A Gly14----Val14 mutant (Gly14 in ras2 is equivalent to Gly12 in mammalian ras proteins) was constructed in ras2 by site-directed mutagenesis. Inducible constructs of ras2 and ras2Val14 were made by coupling the hsp70 promoter to each of the genes. In addition, the endogenous ras2 promoter was placed upstream of the mutated ras2Val14. These constructs were introduced into the germ line of Drosophila to yield transgenic lines by P-element-mediated transformation. A wide variety of developmental disorders were displayed in flies carrying the mutated ras2 gene driven by either the endogenous or the hsp70 promoter. The basal level of transcription of mutated ras from the uninduced hsp70 promoter was sufficient to produce disturbances in the development of several tissue types. In contrast, phenotypic disturbances were not seen with the normal ras2 gene driven by the hsp70 promoter even when induced by heat shock to very high levels of transcription over normal ras2 levels. A subset of the tissues expressing ras2 during development was particularly sensitive to the expression of ras2Val14. The compound eye was found to develop a dorsal-to-ventral 'scar' correlated with the wave of differentiation occurring in the eye imaginal disc at the time of a single brief induction of the hsp70 promoted ras2Val14 construct.

Animals↗
PubMed1987

Developmental expression of Drosophila melanogaster retrovirus-like transposable elements.

We have determined the pattern of temporal expression of several Drosophila retrovirus-like transposable elements. Some of these elements can be grouped into classes whose members show a similar profile of developmental transcription. The members of the 412 class, which includes 412, mdg1, 17.6 and 3S18, are transcribed mainly in the early larval and pupal stages of development, with small differences among the various members. HMS Beagle and Springer constitute another class where RNA accumulation in the larval stages is higher than in pupae and the adult flies accumulate more RNA than any other stage of development. Finally, the transcription of other elements such as copia, 297 and B104 follows a specific and individual pattern distinct from those described above. These results suggest the existence of evolutionary relationships among different transposable elements in Drosophila and the involvement of different cellular genes in the control of their expression.

Animals↗
PubMed1987

Separate regulatory elements are responsible for the complex pattern of tissue-specific and developmental transcription of the yellow locus in Drosophila melanogaster.

DNA sequences involved in the control of the developmental and spatial expression of the yellow locus of Drosophila were identified by phenotypic analysis of germline transformants carrying various in vitro modified yellow genes. Two regions, located between -2873 bp and -1868 bp and between -1868 bp and -700 bp, act as tissue-specific enhancers which respectively regulate yellow transcription in the wings and body of the adult flies. Sequences situated closer to the mRNA cap site, between -225 bp and -91 bp from the start of transcription, are responsible for yellow expression in the denticle belts and mouth parts during larval development. Finally, coloration of the adult bristles is regulated by sequences located in the intron of the yellow gene. These results indicate the existence of several separate DNA elements responsible for the different patterns of temporal and spatial expression of the yellow locus.

Animals↗
PubMed1987

The ovarian, ecdysterone, and heat-shock-responsive promoters of the Drosophila melanogaster hsp27 gene react very differently to perturbations of DNA sequence.

The effect of various types of DNA sequence alterations on the activity of the ovarian, ecdysterone, and heat-shock-responsive promoters of the Drosophila melanogaster hsp27 gene was studied by P element-mediated germ line transformation. Regions of DNA required for proper expression of the gene under these different conditions were identified. Wild-type levels of transcription during oogenesis are dependent on two elements respectively located within a 64-base-pair (bp) fragment in the transcribed untranslated region and between -227 and -958 bp upstream of the transcription start site. This ovarian expression is particularly sensitive to both chromosomal position effects and an increased distance between the distal upstream promoter element and the TATAA homology. The ecdysterone-mediated expression during metamorphosis is dependent on a 145-bp domain including the TATAA box and additional upstream sequences that augment transcription by two- to five-fold. Finally, sequences necessary for heat shock expression are located much further upstream from hsp27 than those previously found for hsp70, although basal expression was correlated with the presence of more proximal heat shock consensus sequences.

Base Sequence↗
PubMed1986

On the molecular mechanism of gypsy-induced mutations at the yellow locus of Drosophila melanogaster.

We determined the nucleotide sequence of genomic DNA corresponding to the yellow gene. The limits of the transcribed region were deduced from sequence analysis of yellow larval and pupal cDNA clones. The yellow transcription unit is simple, composed of two exons which are processed identically in both developmental stages into a mRNA of 1990 bp. The predicted yellow protein has a mol. wt of 60,752 daltons and appears to be a secreted protein having a structural function and not an enzymatic role in pigmentation. We also characterized the spontaneous mutation y2 and a revertant of this allele to investigate the mutagenic effect of the gypsy element inserted into this locus. Our results show that this transposon is inserted at -700 bp and that the y2+ revertant resulted from excision of the gypsy element leaving behind a complete long terminal repeat (LTR). We conclude, therefore, that the gypsy element is neither inserted into a pupal specific intron or regulatory sequence supporting the hypothesis that mutagenesis is a result of transcriptional interference by the gypsy element on the yellow gene.

Amino Acid Sequence↗
PubMed1986

Interactions among the gypsy transposable element and the yellow and the suppressor of hairy-wing loci in Drosophila melanogaster.

We cloned and characterized the yellow locus of Drosophila melanogaster. We also studied its transcription pattern in the suppressible allele y2, which is caused by the insertion of the transposable element gypsy, and the effect of mutations at the unlinked suppressor of Hairy-wing locus on the transcription of yellow RNAs. The gypsy element is transcribed in a temporal fashion that correlates with the pattern of expression of the yellow locus. We propose that the mutational effect of the gypsy element is due to developmentally specific transcriptional interference on yellow transcription. Mutations at the su(Hw) locus reverse this effect by altering the quantitative expression of gypsy.

Animals↗
PubMed1986

The Drosophila melanogaster gypsy transposable element encodes putative gene products homologous to retroviral proteins.

We determined the complete nucleotide sequence of the gypsy element present at the forked locus of Drosophila melanogaster in the f1 allele. The gypsy element shares more homology with vertebrate retroviruses than with the copia element of D. melanogaster or the Ty element of Saccharomyces cerevisiae, both in overall organization and at the DNA sequence level. This transposable element is 7,469 base pairs long and encodes three putative protein products. The long terminal repeats are 482 nucleotides long and contain transcription initiation and termination signals; sequences homologous to the polypurine tract and tRNA primer binding site of retroviruses are located adjacent to the long terminal repeats. The central region of the element contains three different open reading frames. The second one encodes a putative protein which shows extensive amino acid homology to retroviral proteins, including gag-specific protease, reverse transcriptase, and DNA endonuclease.

Amino Acid Sequence↗
PubMed1986

Mutations at the suppressor of forked locus increase the accumulation of gypsy-encoded transcripts in Drosophila melanogaster.

We studied the effect of mutations at the suppressor of forked [su(f)] locus in Drosophila melanogaster on the accumulation of transcripts encoded by the gypsy transposable element. Mutations at this locus do not affect the pattern of developmental expression of gypsy, but they cause an increase in the total amount of gypsy RNA present at different stages of development as compared with wild-type or su(f)/+ flies. These results suggest that the su(f)-encoded products acts as a negative regulator of gypsy expression.

Animals↗
PubMed1985

Forked, gypsys, and suppressors in Drosophila.

We have isolated DNA sequences defining the forked locus of Drosophila. All the alleles belonging to the right pseudoallelic series of M. M. Green map within a 5.4 kb DNA fragment that encodes four transcripts. These RNAs are only expressed in 2-4 day old pupae and their accumulation is affected by mutations at the forked locus. Also, homozygous mutations at the unlinked suppressor loci su(Hw) and su(f) cause the accumulation of these RNAs to return to the normal wild-type levels. The gypsy transposable element, which causes the mutant phenotype in the suppressible forked alleles, encodes a transcript that is maximally expressed in 2-3 day old pupae, the same time in development as transcription of the forked RNAs. We propose a model to explain the mutagenic effect of gypsy based on the influence of promoter elements in the long terminal repeats of the transposon on the transcription from other promoters located nearby.

Alleles↗
PubMed1984

Correct temperature induction and developmental regulation of a cloned heat shock gene transformed into the Drosophila germ line.

We have constructed a size variant of the Drosophila hsp28 gene by deleting 207 base pairs of the protein coding region, beginning 33 base pairs downstream of the ATG protein initiation codon. After transformation of Drosophila melanogaster rosy (ry506) flies with this altered gene, using the P transposable element system, it was found that the transformed gene was regulated correctly both after temperature elevation and during the development of the flies. Levels of the variant mRNA were as high as those of the endogenous hsp28 during all patterns of expression, and the variant mRNA appeared in all cases to be processed correctly and to be as stable as the endogenous mRNA. Nevertheless, the chromosomal locus of the transformed gene did not puff after heat shock, suggesting that normal transcription of the gene does not require puffing of the locus. The deleted hsp28 gene retained the reading frame of the endogenous one, and a protein of the expected molecular weight of 18,500 was made after heat shock at levels comparable to those of the endogenous hsp28.

Amino Acid Sequence↗
PubMed1981

Binding of microtubule protein to DNA and chromatin: possibility of simultaneous linkage of microtubule to nucleic and assembly of the microtubule structure.

Microtubule protein binds to DNA through microtubule associated polypeptides (MAPs). Among MAPs there is one high molecular weight polypeptide (MAP2) which interacts with DNA fundamentally through certain polynucleotide sequences. This interaction is not affected by the presence of histones and other chromosomal proteins. DNA can associate to assembled microtubules and when a determinate DNA/protein ratio is reached the nucleic acid behaves as a microtubule associated molecule. The nucleic acid fragments which preferentially bind to microtubules have been isolated and characterized. These fragments contain DNA regions enriched in repetitive sequences that hybridizes preferentially to the pericentromeric zone of metaphase chromosomes. These results give further support to the model of interaction microtubule-chromosome based upon the mediator function of the microtubule associated proteins.

Animals↗
PubMed1978

Binding of microtubule proteins to DNA: specificity of the interaction.

Tubulin is detected among the DNA-binding proteins when an extract from fibroblasts is chromatographed on DNA-cellulose. Further purification of the colchicine-binding activity shows that purified tubulin from fibroblasts does not bind to DNA. Depolymerized brain microtubule proteins show a high affinity for DNA. The fraction bound is composed of tubulin and microtubule-associated proteins. Experiments with fractionated microtubule proteins indicate that tubulin-free microtubule associated proteins bind to DNA, while tubulin free of microtubule-associated proteins does not. Microtubule-associated proteins bind better to eukaryotic than to phage DNA suggesting a specificity of the interaction.

Animals↗
PubMed1978

Interaction of contractile proteins with DNA.

The interaction of contractile proteins (myosin, actin, tropomyosin and troponin) with DNA was studied in vitro using a nitrocellulose filter binding technique. The data indicate a high affinity of myosin and troponin for DNA, a lesser interaction between DNA and tropomyosin and the absence of binding of actin to DNA. When binding to DNA was detected, the interaction was higher with single-stranded DNA than with RNA or double-stranded DNA, although in some conditions myosin binds equally as well to native as to denatured eukaryotic DNA. Myosin binds better to eukaryotic than to phage native DNA.

Actins↗