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G M Rubin

Publications and source records attributed to G M Rubin.

At least 145 records · Page 8Linked to original sources
PubMed1990

The Drosophila seven-up gene, a member of the steroid receptor gene superfamily, controls photoreceptor cell fates.

The Drosophila seven-up (svp) gene was isolated as a lethal insertion in an "enhancer trap" screen. It is expressed and required in photoreceptor cell precursors R1, R3, R4, and R6 during eye development. The absence of svp+ function causes a transformation of these cells toward an R7 cell fate, as judged by morphology and expression of an R7-specific marker. This transformation depends in part on the sevenless gene product. Our results show that svp is involved in control of cell fate during the generation of neuronal diversity. Molecular analysis of svp reveals that it is a member of the steroid receptor gene superfamily and is likely to be a Drosophila homolog of the human transcription factor COUP.

Amino Acid Sequence↗
PubMed1990

Molecular analysis of no-on-transient A, a gene required for normal vision in Drosophila.

Mutation of the Drosophila melanogaster gene no-on-transient A (nonA) results in reduced visual acuity, behavior abnormalities, and an electrophysiological defect for which the mutant is named. We mapped the nonA gene genetically to a 20 kb interval within the 14C1,2 region of the X chromosome, isolated this chromosomal region, and used P element-mediated transformation to delimit the nonA gene to a 9 kb region. Analysis of cDNA clones indicates that this region encodes alternatively spliced transcripts encoding protein products of approximately 77 kd that differ only in their C-terminal 35 amino acids. Analysis of mutations generated in vitro in this transcription unit confirm that these transcripts are the products of the nonA gene.

Amino Acid Sequence↗
PubMed1990

Comparison of the sevenless genes of Drosophila virilis and Drosophila melanogaster.

The sevenless gene of Drosophila melanogaster encodes a transmembrane tyrosine kinase receptor required for normal eye development. We report here the isolation and DNA sequence analysis of the sevenless gene from Drosophila virilis. The predicted amino acid sequences of the sevenless proteins from these two species, which diverged approximately 60 million years ago, are compared.

Amino Acid Sequence↗
PubMed1990

Structural and functional comparisons of the Drosophila virilis and Drosophila melanogaster rough genes.

We have isolated the homeobox gene rough (ro) from Drosophila virilis. Comparison of the predicted amino acid sequences of the D. melanogaster and D. virilis rough proteins reveals that domains of high conservation, including the homeodomain, are interspersed with highly diverged regions. Stretches of significant sequence conservation are also observed in the 5' promoter region and in the introns. The D. virilis rough gene rescues the rough mutant phenotype and is properly regulated when introduced into the D. melanogaster genome. Thus the rough protein as well as the cis-regulatory elements that ensure proper temporal and spatial regulation are functionally conserved between these Drosophila species.

Amino Acid Sequence↗
PubMed1990

Isolation and expression of scabrous, a gene regulating neurogenesis in Drosophila.

Mutations in the Drosophila scabrous (sca) gene affect eye and bristle development, leading to irregular spacing of ommatidia and bristle duplications in the adult fly. We have cloned the sca gene by P-element tagging. The sca transcription unit is 12 kb and consists of four exons that are joined in a 3.2-kb mRNA. In an enhancer trap screen we have isolated several P[lacZ] insertions close to the sca transcription start site. We have examined the expression pattern of sca by in situ hybridization to sca transcripts, by beta-galactosidase localization in the P[lacZ] lines, and by immunocytochemistry with an anti-sca antiserum. During embryogenesis, sca is expressed in a dynamic pattern associated with neural development. During imaginal development, sca is mainly expressed in the R8 photoreceptor precursor cells in the eye imaginal disc and in sensory organ precursor cells in other discs. In the wing disc, sca expression is coextensive with the anlagen for bristles and is controlled by genes of the achaete-scute complex. Based on its loss-of-function phenotype, expression pattern, and the predicted structure of its product, a secreted peptide with homology to the fibrinogen gene family, we propose that sca encodes a signal involved in lateral inhibition within individual domains of the developing nervous system.

Animals↗
PubMed1990

Analysis of cis-acting requirements of the Rh3 and Rh4 genes reveals a bipartite organization to rhodopsin promoters in Drosophila melanogaster.

The rhodopsin genes of Drosophila melanogaster are expressed in nonoverlapping subsets of photoreceptor cells within the insect visual system. Two of these genes, Rh3 and Rh4, are known to display complementary expression patterns in the UV-sensitive R7 photoreceptor cell population of the compound eye. In addition, we find that Rh3 is expressed in a small group of paired R7 and R8 photoreceptor cells at the dorsal eye margin that are apparently specialized for the detection of polarized light. In this paper we present a detailed characterization of the cis-acting requirements of both Rh3 and Rh4. Promoter deletion series demonstrate that small regulatory regions (less than 300 bp) of both R7 opsin genes contain DNA sequences sufficient to generate their respective expression patterns. Individual cis-acting elements were further identified by oligonucleotide-directed mutagenesis guided by interspecific sequence comparisons. Our results suggest that the Drosophila rhodopsin genes share a simple bipartite promoter structure, whereby the proximal region constitutes a functionally equivalent promoter "core" and the distal region determines cell-type specificity. The expression patterns of several hybrid rhodopsin promoters, in which all or part of the putative core regions have been replaced with the analogous regions of different rhodopsin promoters, provide additional evidence in support of this model.

Animals↗
PubMed1990

The homeo domain protein rough is expressed in a subset of cells in the developing Drosophila eye where it can specify photoreceptor cell subtype.

The Drosophila homeo box gene rough is required in photoreceptor cells R2 and R5 for normal eye development. We show here that rough protein expression is limited to a subset of cells in the developing retina where it is transiently expressed for 30-60 hr. The rough protein is first expressed broadly in the morphogenetic furrow but is rapidly restricted to the R2, R3, R4, and R5 precursor cells. Ubiquitous expression of rough under the control of the hsp70 promoter in third-instar larvae suppresses the initial steps of ommatidial assembly. Structures derived from other imaginal discs are not affected. Ectopic expression of rough in the R7 precursor, through the use of the sevenless promoter, causes this cell to develop into an R1-6 photoreceptor subtype; however, this cell still requires sevenless function for its neural differentiation. Taken together with previous analyses of the rough mutant phenotype, these results suggest that the normal role of rough is to establish the unique cell identity of photoreceptors R2 and R5.

Animals↗
PubMed1989

Effect on eye development of dominant mutations in Drosophila homologue of the EGF receptor.

The compound eye of the adult fruitfly, Drosophila melanogaster, comprises about 800 identical ommatidia, or unit eyes, each containing 20 distinct cells. We have used histological and immunocytochemical methods to study the development of the compound eye in Ellipse (Elp) mutants. In Elp/Elp, most ommatidia do not initiate differentiation. We present genetic evidence that Elp alleles are mutations of the Drosophila homologue of the epidermal growth factor (EGF) receptor, and suggest that activity of the EGF receptor may determine the spacing pattern of ommatidia in the eye.

Animals↗
PubMed1989

cGMP-dependent protein kinase genes in Drosophila.

Two Drosophila genes encoding products related to cGMP-dependent protein kinase have been isolated by cross-hybridization to a Drosophila cAMP-dependent protein kinase catalytic subunit gene. Both genes encode products with putative cGMP binding and kinase domains on the same polypeptide chain, as found for the prototypical bovine lung cGMP-dependent protein kinase. The deduced product of one gene (DG1; cytological position, 21D) is 14% larger than the bovine enzyme and differs substantially in sequence at the amino terminus, the region responsible in the bovine enzyme for dimerization. The second gene (DG2; cytological position, 24A) is transcribed into three major RNA species of different size. The largest (DG2; T1) and smallest (DG2;T3) RNAs encode overlapping polypeptides of similar sequence to the whole length of bovine lung cGMP-dependent protein kinase. The translation product of the third major RNA (DG2;T2) lacks sequences similar to those that constitute the dimerization and kinase inhibitory domains of the bovine enzyme. The percentage amino acid identity between DG1 or DG2 and bovine lung cGMP-dependent protein kinase is 55 and 64%, respectively. A common progenitor of the two cGMP-dependent protein kinase genes, DG1 and DG2, is strongly suggested by the conserved positions of introns in these genes.

Amino Acid Sequence↗
PubMed1989

Ommatidia in the developing Drosophila eye require and can respond to sevenless for only a restricted period.

The sevenless gene encodes a putative cell surface receptor that is expressed in a highly specific and complex pattern in the developing Drosophila eye. We show here that this pattern is not required for the development of either the R7 cell or any other cell in the retina. By transiently expressing sevenless protein, we show that there is only a brief period during eye development when the sevenless protein is required for the formation of the R7 photoreceptor. Our results indicate that the specificity of the cell-cell signaling mediated by the sevenless protein is not achieved by regulating the distribution of the sevenless protein, but rather must reside in some combination of the temporal and spatial distributions of the sevenless ligand or other components of the sevenless signaling pathway.

Animals↗
PubMed1989

Molecular characterization of the Drosophila trp locus: a putative integral membrane protein required for phototransduction.

Recent studies suggest that the fly uses the inositol lipid signaling system for visual excitation and that the Drosophila transient receptor potential (trp) mutation disrupts this process subsequent to the production of IP3. In this paper, we show that trp encodes a novel 1275 amino acid protein with eight putative transmembrane segments. Immunolocalization indicates that the trp protein is expressed predominantly in the rhabdomeric membranes of the photoreceptor cells.

Amino Acid Sequence↗
PubMed1989

Regulation of the complex pattern of sevenless expression in the developing Drosophila eye.

The sevenless gene encodes a cell surface receptor that has protein-tyrosine kinase activity and is expressed in a highly specific and complex pattern in the developing Drosophila eye. We have coupled the sevenless promoter to the reporter gene lacZ and have examined the pattern of beta-galactosidase expression in the developing eyes of transgenic larvae. Our results indicate that the dynamic pattern of sevenless protein expression is regulated transcriptionally. Promoter sequences located 5' of the coding region are insufficient for the wild-type level of gene expression but appear to be able to confer the correct pattern of expression. In contrast, enhancer sequences within the body of the gene can confer both the correct pattern and a normal level of expression on either the sevenless promoter or heterologous promoters. Thus the complex pattern of sevenless expression is redundantly encoded within proximal promoter sequences and enhancer elements internal to the gene but relies on these enhancer sequences for correct quantitative expression.

Animals↗
PubMed1989

Construction, stable transformation, and function of an amber suppressor tRNA gene in Drosophila melanogaster.

Drosophila melanogaster strains with a stably incorporated amber suppressor tRNA gene have been generated. A tRNATyr gene was site specifically mutated to produce an anticodon sequence that recognizes the amber codon and then introduced into Drosophila by using P-element-mediated transformation. Transformants from four integration events were recovered. Two integrations resulted in both male and female sterility, whereas the other two resulted in male sterility but female fertility. Strains derived from the two female-fertile integration events were shown to have a low level of amber-suppressing activity by their ability to suppress an amber mutation in a chloramphenicol acetyltransferase gene.

Animals↗
PubMed1989

Structure and activity of the sevenless protein: a protein tyrosine kinase receptor required for photoreceptor development in Drosophila.

The sevenless gene encodes a putative protein tyrosine kinase receptor that is required for the proper differentiation of the R7 photoreceptor cells of the Drosophila eye. We have expressed the sevenless protein in Drosophila tissue culture cells and studied its synthesis, processing, and activity. Our results show that the sevenless protein possesses protein tyrosine kinase activity. The protein is first synthesized as a 280-kDa glycoprotein precursor that is subsequently cleaved into 220-kDa amino-terminal and 58-kDa carboxyl-terminal subunits that remain associated by noncovalent interactions. The 220-kDa subunit is glycosylated and contains most of the extracellular portion of the protein, and the 58-kDa subunit is composed of a small portion of the extracellular sequences and the intracellular protein tyrosine kinase domain. This complex is subsequently cleaved into either 49- or 48-kDa carboxyl-terminal fragments with concomitant degradation of the rest of the protein.

Animals↗
PubMed1989

Definition of cis-acting elements regulating expression of the Drosophila melanogaster ninaE opsin gene by oligonucleotide-directed mutagenesis.

We have analyzed the cis-acting regulatory sequences of the Rh1 (ninaE) gene in Drosophila melanogaster by P-element-mediated germline transformation of indicator genes transcribed from mutant ninaE promoter sequences. We have previously shown that a 200-bp region extending from -120 to +67 relative to the transcription start site is sufficient to obtain eye-specific expression from the ninaE promoter. In the present study, 22 different 4-13-bp sequences in the -120/+67 promoter region were altered by oligonucleotide-directed mutagenesis. Several of these sequences were found to be required for proper promoter function; two of these are conserved in the promoter of the homologous gene isolated from the related species Drosophila virilis. Alteration of a conserved 9-bp sequence results in aberrant, low level expression in the body. Alteration of a separate 11-bp sequence, found in the promoter regions of several photoreceptor-specific genes of Drosophila, results in an approximately 15-fold reduction in promoter efficiency but without apparent alteration of tissue-specificity. A protein factor capable of interacting with this 11-bp sequence has been detected by DNaseI footprinting in embryonic nuclear extracts. Finally, we have further characterized two separable enhancer sequences previously shown to be required for normal levels of expression from this promoter.

Animals↗
PubMed1989

Analysis of the cis-acting requirements for germ-line-specific splicing of the P-element ORF2-ORF3 intron.

P-element transposition is limited to the germ line because the element's third intron is only spliced in germ line cells. We show that a 240-bp fragment containing this 190-bp intron can confer germ line specificity when placed in the context of another gene. We find that the cis-acting regulatory sequences required for germ line regulation map near to, but not at, the 5' or 3' splice junctions.

Animals↗