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At least 217 records · Page 12Linked to original sources
PubMed1993

A positive role in differentiation for the male doublesex protein of Drosophila.

The doublesex (dsx) locus encodes male-specific and female-specific polypeptides that are essential for the proper differentiation of sexually dimorphic somatic features of Drosophila melanogaster. Ectopic expression of the male-specific dsx polypeptide was obtained by P-element-mediated transformation of flies with a construct bearing a fusion between the hsp70 heat shock promoter and dsx male-specific cDNA sequences. Heat shock-induced expression of the male cDNA in either sex resulted in three novel phenotypes: transformation of bristles on all legs toward a sex comb-like morphology, pigmentation of dorsal spinules and ventral setae in third-instar larvae, and lethality. These results were not predicted by previous models of dsx function, and provide evidence that the role of the male dsx protein includes activation of some aspects of male differentiation as well as repression of female differentiation.

Animals↗
PubMed1995

Neoplastic transformation and aberrant cell-cell interactions in genetic mosaics of lethal(2)giant larvae (lgl), a tumor suppressor gene of Drosophila.

Homozygosity for lethal(2)giant larvae (lgl), a mutation in a tumor suppressor gene of Drosophila, induces neoplasia of the imaginal discs. To explore the developmental capacities of lgl mutant cells, we have investigated their growth and differentiation in genetic mosaics. Adult wings mosaic for lgl displayed abnormal growth and differentiation of the lgl mutant and neighboring wild-type cells, suggesting aberrant cell-cell interactions during development. lgl mutant clones also straddled the anteroposterior boundary of the wing imaginal disc, apparently due to failure of the cells of the anterior and the posterior compartment to segregate at the boundary. To further test if anteroposterior compartmentalization takes place in the neoplastic imaginal discs of lgl mutant larvae, we studied the expression of an engrailed (en)-specific lacZ reporter gene during progressive stages of their tumorous growth. Our results show that en is activated in the posterior compartments of the neoplastic imaginal discs. However, during later stages of tumorous overgrowth, the en-expressing and nonexpressing cells appear to show extensive intermixing. These observations suggest that neoplastic transformation of imaginal discs involves loss of their normal cell-cell interactions and signaling.

Animals↗
PubMed2001

Isolation of mutations that disrupt cooperative DNA binding by the Drosophila bicoid protein.

Cooperative DNA binding is thought to contribute to the ability of the Drosophila melanogaster protein, Bicoid, to stimulate transcription of target genes in precise sub-domains within the embryo. As a first step toward testing this idea, we devised a genetic screen to isolate mutations in Bicoid that specifically disrupt cooperative interactions, but do not disrupt DNA recognition or transcription activation. The screen was carried out in Saccharomyces cerevisiae and 12 cooperativity mutants were identified. The mutations map across most of the Bicoid protein, with some located within the DNA-binding domain (homeodomain). Four homeodomain mutants were characterized in yeast and shown to activate a single-site reporter gene to levels comparable to that of wild-type, indicating that DNA binding per se is not affected. However, these mutants failed to show cooperative coupling between high and low-affinity sites, and showed reduced activation of a reporter gene carrying a natural Drosophila enhancer. Homology modeling indicated that none of the four mutations is in residues that contact DNA. Instead, these residues are likely to interact with other DNA-bound Bicoid monomers or other parts of the Bicoid protein. In vitro, the isolated homeodomains did not show strong cooperativity defects, supporting the idea that other regions of Bicoid are also important for cooperativity. This study describes the first systematic screen to identify cooperativity mutations in a eukaryotic DNA-binding protein.

Allosteric Regulation↗
PubMed1996

Regulation, function and potential origin of the Drosophila gene spalt adjacent, which encodes a secreted protein expressed in the early embryo.

During early embryogenesis of Drosophila the spatial and temporal expression patterns of the region-specific homeotic gene spalt (sal) and the neighbouring gene spalt adjacent (sala) extensively overlap. We show that the initial expression patterns of the two genes in the blastoderm also have identical genetic controls. However, while sal encodes a transcription factor, sala encodes a precursor protein from which a functional signal peptide is cleaved off to generate the secreted sala protein. Ectopic expression or absence of sala protein does not affect embryonic development, adult viability or fertility. In addition to sal and sala, we identified a third gene nearby, termed spalt related (salr), which shares coding sequence similarity and a late embryonic expression pattern with sal, but lacks the early expression domains that are shared by sal and sala. These results suggest that the three genes and their present cis-regulatory regions arose through a chromosomal rearrangement involving local duplication and transposition events in the 32F/33A region on the left arm of the second chromosome.

Amino Acid Sequence↗
PubMed1999

Absence of protein polymorphism in the Ras genes of Drosophila melanogaster.

Sequence analysis of 27 alleles of each of the three Ras-related genes in Drosophila melanogaster indicates that they all have low levels of polymorphism but may experience slightly different evolutionary pressures. No amino acid replacement substitutions were indicated in any of the sequences, or in the sibling species D. simulans and D. mauritiana. The Dras1 gene, which is the major ras homologue in Drosophila, has less within-species variation in D. melanogaster relative to the amount of divergence from the sibling species than does Dras2, although the contrast was not significant by the HKA test. Dras2 appears to be maintaining two classes of haplotype in D. melanogaster, one of which is closer to the alleles observed in the sibling species, suggesting that this is not likely to be a pseudogene despite the absence of a mutant phenotype. Although differences in level of expression may affect the function of the genes, it is concluded that genetic variation in the Ras signal transduction pathways cannot be attributed to catalytic variation in the Ras proteins.

Alleles↗
PubMed2001

Differentially expressed regulatory genes in honey bee caste development.

In the honey bee, an eminently fertile queen with up to 200 ovarioles per ovary monopolizes colony level reproduction. In contrast, worker bees have only few ovarioles and are essentially sterile. This phenotype divergence is a result of caste-specifically modulated juvenile hormone and ecdysteroid titers in larval development. In this study we employed a differential-display reverse transcription (DDRT)-PCR protocol to detect ecdysteroid-regulated gene expression during a critical phase of caste development. We identified a Ftz-F1 homolog and a Cut-like transcript. Ftz-F1 could be a putative element of the metamorphic ecdysone response cascade of bees, whereas Cut-like proteins are described as transcription factors involved in maintaining cellular differentiation states. The downregulation of both factors can be interpreted as steps in the metamorphic degradation of ovarioles in worker-bee ovaries.

Amino Acid Sequence↗
PubMed1993

The segment polarity gene hedgehog is required for progression of the morphogenetic furrow in the developing Drosophila eye.

Cell-type specification in the Drosophila compound eye begins at the morphogenetic furrow. The furrow sweeps across the developing eye epithelium and is coincident with four classes of cellular events: coordinated changes in cell shape, changes in gene expression, synchronization of the cell cycle, and the specification of a regular array of ommatidial founder cells. The molecular mechanisms that induce these events in the developing eye have hitherto been unknown. We identify here a gene specifically required for furrow progression, hedgehog (hh). We show that hh expression posterior to the morphogenetic furrow is continuously required for its progression. We propose that forward diffusion of hh protein induces anterior cells to enter the furrow.

Animals↗
PubMed1995

Use of juvenile hormone esterase as a novel reporter enzyme in the baculovirus expression system.

Juvenile hormone esterase (JHE) has a number of characteristics favorable for use as a reporter enzyme. It is extremely stable under a variety of adverse conditions including in organic solvent. JHE is easily detected by a rapid and sensitive colorimetric assay, and detection is facilitated by export of the enzyme from the cell. Its use is illustrated in the baculovirus expression system by promoter studies and evaluation of culture conditions necessary for optimal production of recombinant proteins. From this, it was found that protein yields were greater for Autographa californica nuclear polyhedrosis virus (AcNPV) expression vectors using the basic protein promoter to drive production of JHE than for the p10 protein or polyhedrin promoters. This has significant implications for current baculovirus expression methodologies. The effect of multiplicity of infection on protein yield was found to be insignificant between 0.1 and 10 for expression under the basic protein promoter. Yields of JHE were about 40% higher from the cell line Tn5B1-4 ('High Five') relative to the Sf21 cell line under optimized conditions for each cell line, with maximum yields obtained at 2-3 days, and 3-5 days post-infection for the two cell lines respectively when cultured in ExCell 401 medium. The presence of fetal calf serum in the cell culture medium enhanced protein yields from both cell lines. These studies demonstrate the use of JHE as a reporter enzyme for optimizing high yields of protein from the baculovirus expression system. JHE also has a potential application as a reporter enzyme in other eukaryotic systems. The advantages and use of JHE over other reporter enzyme are discussed.

Animals↗
PubMed1994

The cold-sensitive period for frizzled in the development of wing hair polarity ends prior to the start of hair morphogenesis.

The function of the frizzled (fz) gene is essential for the development of the normal pattern of hairs on the Drosophila wing. In the absence of fz function hairs develop, but they display an abnormal polarity. Mutations in fz result in an altered subcellular location for the assembly of the F-actin filled prehair that becomes the adult cuticular hair. This observation led to the suggestion that fz and other tissue polarity genes form a regulatory pathway that controls the initiation of prehairs. We have isolated a cold-sensitive fz allele and found that the cold-sensitive period for fz in the pupal wing starts in the early pupae and ends prior to the first sign of prehair morphogenesis. This cold-sensitive mutation is due to a missense mutation in a putative transmembrane domain. Western blot analysis shows that the accumulation of the mutant protein is not cold sensitive, consistent with the supposition that it is the activity of the mutant protein that is cold sensitive. Our data argue that fz has a regulatory function in specifying where the prehair forms, but no role in the actual morphogenesis of the prehair.

Actins↗
PubMed1996

The dorsoventral regulatory gene cassette spätzle/Toll/cactus controls the potent antifungal response in Drosophila adults.

The cytokine-induced activation cascade of NF-kappaB in mammals and the activation of the morphogen dorsal in Drosophila embryos show striking structural and functional similarities (Toll/IL-1, Cactus/I-kappaB, and dorsal/NF-kappaB). Here we demonstrate that these parallels extend to the immune response of Drosophila. In particular, the intracellular components of the dorsoventral signaling pathway (except for dorsal) and the extracellular Toll ligand, spätzle, control expression of the antifungal peptide gene drosomycin in adults. We also show that mutations in the Toll signaling pathway dramatically reduce survival after fungal infection. Antibacterial genes are induced either by a distinct pathway involving the immune deficiency gene (imd) or by combined activation of both imd and dorsoventral pathways.

Animals↗
PubMed1996

A cyclin-dependent kinase inhibitor, Dacapo, is necessary for timely exit from the cell cycle during Drosophila embryogenesis.

In a screen for genes that interact with the Rap1 GTPase, we have identified a Drosophila gene, dacapo (dap), which is a member of the p21/p27 family of cdk inhibitors. Unlike mammalian cdk inhibitors studied to date, dap is essential for normal embryonic development. Dacapo inhibits cyclin-cdk activity in vitro. Overexpressing dap during eye development interferes with cell cycle progression and interacts genetically with the retinoblastoma homolog (Rbf) and cyclin E. dap expression in embryos parallels the exit of cells from the cell cycle. dap mutant embryos delay the normal cell cycle exit during development; many cells complete an additional cycle and subsequently become quiescent. Thus, dap functions during embryogenesis to achieve a precisely timed exit from the cell cycle.

Amino Acid Sequence↗
PubMed1997

Unique gene organization: alternative splicing in Drosophila produces two structurally unrelated proteins.

The Ub80 gene in eukaryotes produces a ubiquitin fusion protein in which ubiquitin is fused in frame to a tail protein (Redman and Rechsteiner, 1988; Finley et al., 1989; Barrio et al., 1994). The tail protein is incorporated into the ribosome, and ubiquitin is thought to act as a chaperone. The DUb80 gene of Drosophila melanogaster was cloned by Barrio et al. (1994) and contains a 5'-untranslated exon, followed by a large intron and then the first coding exon. We report that the large intron of DUb80 contains an open reading frame, which produces a 259-aa protein (IP259) that is conserved in eukaryotes from yeast to mammals. Transcription of the DUb80 and IP259 mRNAs begins at the same start sites. However, alternate splicing of the primary transcript produces two structurally unrelated proteins. This is the second reported instance of two structurally unrelated proteins being produced via alternate splicing, suggesting that this form of genomic organization may be more common than previously thought.

Amino Acid Sequence↗
PubMed2000

Role of AWD/nucleoside diphosphate kinase in Drosophila development.

The abnormal wing discs gene of Drosophila encodes a soluble protein with nucleoside diphosphate kinase activity. This enzymic activity is necessary for the biological function of the abnormal wing discs gene product. Complete loss of function, i.e., null, mutations cause lethality after the larval stage. Most larval organs in such null mutant larvae appear to be normal, but the imaginal discs are small and incapable of normal differentiation. Killer-of-prune is a neomorphic mutation in the abnormal wing discs gene. It causes dominant lethality in larvae that lack prune gene activity. The Killer-of-prune mutant protein may have altered substrate specificity. Null mutant larvae have a low level of nucleoside diphosphate kinase activity. This suggests that there may be additional Drosophila genes that encode proteins with nucleoside dipthosphate kinase activity. Candidate genes have been found in the Drosophila genome.

Animals↗
PubMed1996

Focus on butterfly eyespot development.

The beautiful eyespot patterns on the wings of butterflies help them to evade predators, but they also present biologists with a unique opportunity to investigate the reciprocal interactions of development and evolution.

Adaptation, Biological↗
PubMed1996

Development, plasticity and evolution of butterfly eyespot patterns.

The developmental and genetic bases for the formation, plasticity and diversity of eyespot patterns in butterflies are examined. Eyespot pattern mutants, regulatory gene expression, and transplants of the eyespot developmental organizer demonstrate that eyespot position, number, size and colour are determined progressively in a developmental pathway largely uncoupled from those regulating other wing-pattern elements and body structures. Species comparisons and selection experiments suggest that the evolution of eyespot patterns can occur rapidly through modulation of different stages of this pathway, and requires only single, or very few, changes in regulatory genes.

Adaptation, Biological↗
PubMed2002

Tsc tumour suppressor proteins antagonize amino-acid-TOR signalling.

Target of Rapamycin (TOR) mediates a signalling pathway that couples amino acid availability to S6 kinase (S6K) activation, translational initiation and cell growth. Here, we show that tuberous sclerosis 1 (Tsc1) and Tsc2, tumour suppressors that are responsible for the tuberous sclerosis syndrome, antagonize this amino acid-TOR signalling pathway. We show that Tsc1 and Tsc2 can physically associate with TOR and function upstream of TOR genetically. In Drosophila melanogaster and mammalian cells, loss of Tsc1 and Tsc2 results in a TOR-dependent increase of S6K activity. Furthermore, although S6K is normally inactivated in animal cells in response to amino acid starvation, loss of Tsc1-Tsc2 renders cells resistant to amino acid starvation. We propose that the Tsc1-Tsc2 complex antagonizes the TOR-mediated response to amino acid availability. Our studies identify Tsc1 and Tsc2 as regulators of the amino acid-TOR pathway and provide a new paradigm for how proteins involved in nutrient sensing function as tumour suppressors.

Amino Acids↗
PubMed2000

Mutations in the beta-propeller domain of the Drosophila brain tumor (brat) protein induce neoplasm in the larval brain.

Inactivation of both alleles of the fruit fly D. melanogaster brain tumor (brat) gene results in the production of a tumor-like neoplasm in the larval brain, and lethality in the larval third instar and pupal stages. We cloned the brat gene from a transposon-tagged allele and identified its gene product. brat encodes for an 1037 amino acid protein with an N-terminal B-boxl zinc finger followed by a B-box2 zinc finger, a coiled-coil domain, and a C-terminal beta-propeller domain with six blades. All these motifs are known to mediate protein-protein interactions. Sequence analysis of four brat alleles revealed that all of them are mutated at the beta-propeller domain. The clustering of mutations in this domain strongly suggests that it has a crucial role in the normal function of Brat, and defines a novel protein motif involved in tumor suppression activity. The brat gene is expressed in the embryonic central and peripheral nervous systems including the embryonic brain. In third instar larva brat expression was detected in the larval central nervous system including the brain and the ventral ganglion, in two glands - the ring gland and the salivary gland, and in parts of the foregut - the gastric caecae and the proventriculus. A second brat-like gene was found in D. melanogaster, and homologs were identified in the nematode, mouse, rat, and human. Accumulated data suggests that Brat may regulate proliferation and differentiation by secretion/transport-mediated processes.

Amino Acid Sequence↗
PubMed2001

Control of Drosophila perineurial glial growth by interacting neurotransmitter-mediated signaling pathways.

Drosophila peripheral nerves, similar structurally to the peripheral nerves of mammals, comprise a layer of axons and inner glia, surrounded by an outer perineurial glial layer. Although it is well established that intercellular communication occurs among cells within peripheral nerves, the signaling pathways used and the effects of this signaling on nerve structure and function remain incompletely understood. Here we demonstrate with genetic methods that the Drosophila peripheral nerve is a favorable system for the study of intercellular signaling. We show that growth of the perineurial glia is controlled by interactions among five genes: ine, which encodes a putative neurotransmitter transporter; eag, which encodes a potassium channel; push, which encodes a large, Zn(2+)-finger-containing protein; amn, which encodes a putative neuropeptide related to the pituitary adenylate cyclase activator peptide; and NF1, the Drosophila ortholog of the human gene responsible for type 1 neurofibromatosis. In other Drosophila systems, push and NF1 are required for signaling pathways mediated by Amn or the pituitary adenylate cyclase activator peptide. Our results support a model in which the Amn neuropeptide, acting through Push and NF1, inhibits perineurial glial growth, whereas the substrate neurotransmitter of Ine promotes perineurial glial growth. Defective intercellular signaling within peripheral nerves might underlie the formation of neurofibromas, the hallmark of neurofibromatosis.

Animals↗