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PubMed1997

About the origin of retroviruses and the co-evolution of the gypsy retrovirus with the Drosophila flamenco host gene.

The gypsy element of Drosophila melanogaster is the first retrovirus identified so far in invertebrates. According to phylogenetic data, gypsy belongs to the same group as the Ty3 class of LTR-retrotransposons, which suggests that retroviruses evolved from this kind of retroelements before the radiation of vertebrates. There are other invertebrate retroelements that are also likely to be endogenous retroviruses because they share with gypsy some structural and functional retroviral-like characteristics. Gypsy is controlled by a Drosophila gene called flamenco, the restrictive alleles of which maintain the retrovirus in a repressed state. In permissive strains, functional gypsy elements transpose at high frequency and produce infective particles. Defective gypsy proviruses located in pericentromeric heterochromatin of all strains seem to be very old components of the genome of Drosophila melanogaster, which indicates that gypsy invaded this species, or an ancestor, a long time ago. At that time, Drosophila melanogaster presumably contained permissive alleles of the flamenco gene. One can imagine that the species survived to the increase of genetic load caused by the retroviral invasion because restrictive alleles of flamenco were selected. The characterization of a retrovirus in Drosophila, one of the most advanced model organisms for molecular genetics, provides us with an exceptional clue to study how a species can resist a retroviral invasion.

Animals↗
PubMed1997

A simulation of P element horizontal transfer in Drosophila.

Experimental data suggest that the P transposable element has invaded the Drosophila melanogaster genome after a horizontal transfer from the phylogenetically distant species Drosophila willistoni. The differences between P element phylogeny and that of the Drosophila genus could in part be explained by horizontal transfers. In vivo experiments show that P elements are able to transpose in the genomes of other Drosophila species. This suggests that horizontal transmission of P elements could have taken place in many species of this genus. The regulation, transposition, and deleterious effects of the P element in D. melanogaster were formalized and integrated in a global model to produce a simulation program that simulates a P element invasion. The simulations show that our knowledge of the P element in D. melanogaster can explain its behavior in the Drosophila genus. The equilibrium state of the invaded population of a new species depends on its ability to repair damage caused by P element activity. If repair is efficient, the equilibrium state tends to be of the P type state, in which case the element could subsequently invade other populations of the species. Conversely, the equilibrium state is of the M' type state when the ability to repair damage is low. The invasion of the P element into other populations of this new species can then only occur by genetic drift and it is likely to be lost. The success of a P element invasion into a new species thus greatly depends on its ability to produce dysgenic crosses.

Animals↗
PubMed1998

Genetic and hormonal control of cytosolic malate dehydrogenase activity in Drosophila melanogaster.

The activity of Drosophila cytosolic malate dehydrogenase (MDH; EC 1.1.1.40), a key enzyme in the biosynthesis of lipids, was found to be regulated by both the sesquiterpenoid juvenile hormone (JH), and the steroid hormone ecdysone. The responsiveness of MDH to JH largely depended on the developmental stage or endogenous titre of ecdysone. During the early through middle period of the last larval instar, when ecdysone levels are low, MDH responded to JH rapidly by increasing activity, while little or no response was measured in mature larvae (postfeeding stage) and fresh pupae when the endogenous pulse of ecdysone is high. Activity of MDH in ecd1 and su(f)ts67g, two ecdysone-deficient mutants of Drosophila, was increased when compared to wild type controls, and was also sensitive to administration of JH. The differences in MDH activity between ecd1 and su(f)ts67g were negligible indicating a substantial role of ecdysone in the enzyme regulation and minimal or no effect of their genetic backgrounds. Accordingly, another Drosophila mutant, ap4 which is naturally deficient in JH production, displayed significantly reduced activity of MDH in heterozygotic combination, and almost undetectable MDH activity in null homozygote adults. The ap4 phenotype was more strongly manifested in the adult stage than in larvae which showed 7-times lower requirements for JH titre. In addition, high/low sucrose diet fed to wild types or mutants affected the activity of larval MDH, but the enzyme remained sensitive to administration of JH. These results corroborate those described for mammals and provide the first evidence that Drosophila MDH might be under differential hormonal and nutritional control. These data will serve as a basis for further molecular characterization of the Drosophila MDH gene and its regulation.

Animals↗
PubMed1998

Microinjected antisera against ductin affect gastrulation in Drosophila melanogaster.

Ductin is a putative connexon-forming protein in gap junctions of arthropods. To analyze the role of gap-junction mediated cell-cell communication during Drosophila embryogenesis, we used two different polyclonal anti-ductin sera. One antiserum was directed against ductin isolated from gap junctions of the lobster Nephrops whilst the other was raised against a nonapeptide at the N-terminus of ductin from Drosophila. Both antisera were found to inhibit, when microinjected into Drosophila ovarian follicles, the intercellular exchange of fluorescent tracer molecules between oocyte and follicle epithelium. This result indicates that Drosophila ductin plays a decisive role in gap-junctional communication and confirms the cytoplasmic location of the ductin N-terminus in gap junctions. On immunofluorescence preparations and immunoblots, the anti-ductin sera specifically recognized ovarian as well as embryonic antigens. Following microinjections of the antisera into embryos prior to gastrulation, significantly reduced rates of hatching larvae were obtained. Moreover, microinjections into the mid-ventral region of the embryos resulted in specific ventral defects that depended on the concentration of the ductin antibodies. In particular, larvae with ventral holes in their cuticles occurred with high frequency. During gastrulation, antiserum-injected embryos often developed defects in the middle region of their ventral furrow. Here, mesodermal cells failed to invaginate correctly and, thus, no cuticle was formed. We conclude that, during Drosophila embryogenesis, gap-junctional communication is required for epithelial integrity and morphogenetic events.

Animals↗
PubMedDate not supplied

[Do viruses participate in the origin of group outbursts of gene mutations occurring in natural Drosophila populations?].

It has been experimentally shown that different viruses apathogenic for Drosophila and unable to reproduce in it, and also highly polymerized DNA isolated from these viruses, act as effective mutagens when injected into the hemolymph of this insect. They induce numerous lethal and visible stable gene mutations. Some of them can be present in Drosophila natural populations. But these virus induced gene mutations or those induced by virus DNA never take part in the origin of group outbursts of gene mutation which are observed in Drosophila melanogaster natural populations. These mutability outbursts include only gene mutations induced by transpositions of mobile genetic elements. These mutations are usually unstable: they are apt to be converted to other allelic mutant states which are phenotypically differently expressed. Viruses which are endemic for Drosophila melanogaster and can reproduce in it take no part in the outbursts of gene mutations. They can only induce a small number of lethal and visible gene mutations which are stable and arise very infrequently. It seems probable that group outbursts of gene mutations in natural populations of Drosophila melanogaster are a result of various environmental stresses which cause a rise in the frequency of transpositions of mobile genetic elements, this leading to the mutability outbursts of gene mutations.

Animals↗
PubMed2001

Painting of fourth, a chromosome-specific protein in Drosophila.

Chromosome-specific gene regulation is known thus far only as a mechanism to equalize the transcriptional activity of the single male X chromosome with that of the two female X chromosomes. In Drosophila melanogaster, a complex including the five Male-Specific Lethal (MSL) proteins, "paints" the male X chromosome, mediating its hypertranscription. Here, with the molecular cloning of Painting of fourth (Pof), we describe a previously uncharacterized gene encoding a chromosome-specific protein in Drosophila. Unlike the MSL proteins, POF paints an autosome, the fourth chromosome of Drosophila melanogaster. Chromosome translocation analysis shows that the binding depends on an initiation site in the proximal region of chromosome 4 and spreads in cis to involve the entire chromosome. The spreading depends on sequences or structures specific to chromosome 4 and cannot extend to parts of other chromosomes translocated to the fourth. Spreading can also occur in trans to a paired homologue that lacks the initiation region. In the related species Drosophila busckii, POF paints the entire X chromosome exclusively in males, suggesting relationships between the fourth chromosome and the X and between POF complexes and dosage-compensation complexes.

Amino Acid Sequence↗
PubMed2024

Misregulation of bromotyrosine compromises fertility in male Drosophila.

Biological regulation often depends on reversible reactions such as phosphorylation, acylation, methylation, and glycosylation, but rarely halogenation. A notable exception is the iodination and deiodination of thyroid hormones. Here, we report detection of bromotyrosine and its subsequent debromination during Drosophila spermatogenesis. Bromotyrosine is not evident when Drosophila express a native flavin-dependent dehalogenase that is homologous to the enzyme responsible for iodide salvage from iodotyrosine in mammals. Deletion or suppression of the dehalogenase-encoding condet (cdt) gene in Drosophila allows bromotyrosine to accumulate with no detectable chloro- or iodotyrosine. The presence of bromotyrosine in the cdt mutant males disrupts sperm individualization and results in decreased fertility. Transgenic expression of the cdt gene in late-staged germ cells rescues this defect and enhances tolerance of male flies to bromotyrosine. These results are consistent with reversible halogenation affecting Drosophila spermatogenesis in a process that had previously eluded metabolomic, proteomic, and genomic analyses.

Animals↗
PubMed1994

Drosophila mode of metamerization in the embryogenesis of the lepidopteran insect Manduca sexta.

Insect embryos have been classified as intermediate- and short-germ embryos, in which posterior segments are thought to be generated sequentially from an uncommitted growth zone, or as long-term embryos, such as Drosophila melanogaster, which develop primordia for all segments simultaneously. In Drosophila the coordinated activities among a three-tiered cascade of zygotic segmentation genes subdivide the embryo into progressively smaller units along the anterior-posterior axis. The mode of pattern specification in lepidopteran embryos has not been determined, although on morphological grounds they have been characterized as intermediate-germ insects. We have cloned orthologues of Drosophila segmentation genes from the tobacco hawkmoth Manduca sexta and have found that the blastoderm expression patterns of these genes show a molecular prepatterning typical of Drosophila. Thus, successive segment formation in Manduca embryos may not be due to sequential addition but rather may be the consequence of a lateral compression of the embryo proceeding in an anterior-to-posterior progression. These data challenge the view that the classification of insect development according to morphological criteria can serve as a reliable indicator of the molecular mechanisms underlying segmentation.

Amino Acid Sequence↗
PubMed1997

Twist-mediated activation of the NK-4 homeobox gene in the visceral mesoderm of Drosophila requires two distinct clusters of E-box regulatory elements.

NK-4, also called msh2 and tinman, encodes a homeodomain transcription factor that is required for the development of the dorsal mesoderm and its derivatives in the Drosophila embryo. Genetic analyses indicate that NK-4 resides downstream of the mesodermal determinant twist, which encodes a basic helix-loop-helix-type transcription factor. However, the regulation of NK-4 by twist remains poorly understood. Using expression assays in cultured cells and transgenic flies, we show that two distinct clusters of E-box regulatory sequences, present upstream of the NK-4 gene, mediate NK-4 expression in the visceral mesoderm. These elements are conserved between the Drosophila melanogaster and Drosophila virilis NK-4 genes and serve as binding sites for Twist (E1 cluster) and NK-4 (E2 cluster) proteins. In cultured cells, Twist and NK-4 binding results in activation of NK-4 gene expression. In transgenic animals, the E1 and E2 clusters are functionally connected, and both elements are required for NK-4 activation in cells of the visceral mesoderm and also for NK-4 repression in cells of the somatic musculature. These results demonstrate that NK-4 is a direct transcriptional target for Twist and its own gene product in visceral mesodermal cells, supporting the idea that twist and NK-4 function in the subdivision of the mesoderm during Drosophila embryogenesis.

Animals↗
PubMedDate not supplied

Species-specific patterns of sexual dimorphism in the expression of fruitless protein, a neural musculinizing factor in Drosophila.

In Drosophila melanogaster, male-specific forms of the fruitless (fru) gene product, mFru protein, function as a neural sex-determination factors that directs the development of at least two male characteristics, namely courtship and mating behavior and the formation of the muscle of Lawrence (MOL). In D. melanogaster, the male-specific expression of Fru protein in motoneurons is responsible for the male-limited induction of the MOL by such neurons. Although no Drosophila species whose females have the MOL are known, there are many Drosophila species whose males lack the MOL. We performed immunohistochemical staining of the central nervous system (CNS) from 9 Drosophila species to determine whether the mFru expression profile is different between MOL-present and MOL-absent species. In 8 of the 9 species, Fru protein expression in the CNS is strictly male-specific, regardless of the presence or absence of the MOL. The sole exception is D. suzukii, in which females express the Fru protein though less extensively than males do: Fru expression in the CNS of female D. suzukii is restricted to the lamina and ventral ganglia. Expression of Fru protein in the lamina is observed in males of D. virilis and in both sexes of D. suzukii, but not in males and females of the 7 other species. These results indicate that sexually dimorphic expression of the Fru protein has been subjected to species-specific modulation during evolution.

Animals↗
PubMed1998

Rate variation of DNA sequence evolution in the Drosophila lineages.

Rate constancy of DNA sequence evolution was examined for three species of Drosophila, using two samples: the published sequences of eight genes from regions of the normal recombination rates and new data of the four AS-C (ac, sc, l'sc and ase) and ci genes. The AS-C and ci genes were chosen because these genes are located in the regions of very reduced recombination in Drosophila melanogaster and their locations remain unchanged throughout the entire lineages involved, yielding less effect of ancestral polymorphism in the study of rate constancy. The synonymous substitution pattern of the three lineages was found to be erratic in both samples. The dispersion index for replacement substitution was relatively high for the per, G6pd and ac genes. A significant heterogeneity was found in the number of synonymous substitutions in the three lineages between the two samples of genes with different recombination rates. This is partly due to a lack of the lineage effect in the D. melanogaster and Drosophila simulans lineages in the AS-C and ci genes in contrast to Akashi's observation of genes in regions of normal recombination. The higher codon bias in Drosophila yakuba as compared with D. melanogaster and D. simulans was observed in the four AS-C genes, which suggests change(s) in action of natural selection involved in codon usage on these genes. Fluctuating selection intensity may also be responsible for the observed locus-lineage interaction effects in synonymous substitution.

Animals↗
PubMed1998

Microevolutionary divergence pattern of the segmentation gene hunchback in Drosophila.

To study the microevolutionary processes shaping the evolution of the segmentation gene hunchback (hb) from Drosophila melanogaster, we cloned and sequenced the gene from 12 isofemale lines representing wild-type populations of D. melanogaster, as well as from the closely related species Drosophila sechellia, Drosophila orena, and Drosophila yakuba. We find a relatively low degree of sequence variation in D. melanogaster (theta = 0.0017), which is, however, consistent with its chromosomal location in a region of low recombination. Tests of neutrality do not reject a neutral-evolution model for the whole region. However, pairwise tests with different subregions indicate that there is a relative excess of polymorphic sites in the leader and the intron. Codon usage pattern analysis shows a particularly biased codon usage in the highly conserved regions, which is in line with the hypothesis that selection on translational accuracy is the driving force behind such a bias. A comparison of the expression pattern of hb in different sibling species of D. melanogaster reveals some regulatory changes in D. yakuba, which could be interpreted as changes in the timing of secondary expression domains.

Animals↗
PubMed1998

Functional analysis of eve stripe 2 enhancer evolution in Drosophila: rules governing conservation and change.

Experimental investigations of eukaryotic enhancers suggest that multiple binding sites and trans-acting regulatory factors are often required for wild-type enhancer function. Genetic analysis of the stripe 2 enhancer of even-skipped (eve), an important developmental gene in Drosophila, provides support for this view. Given the importance of even-skipped expression in early Drosophila development, it might be predicted that many structural features of the stripe 2 enhancer will be evolutionarily conserved, including the DNA sequences of protein binding sites and the spacing between them. To test this hypothesis, we compared sequences of the stripe 2 enhancer between four species of Drosophila: D. melanogaster, D. yakuba, D. erecta and D. pseudoobscura. Our analysis revealed a large number of nucleotide substitutions in regulatory protein binding sites for bicoid, hunchback, Kruppel and giant, as well as a systematic change in the size of the enhancer. Some of the binding sites in D. melanogaster are either absent or modified in other species. One functionally important bicoid-binding site in D. melanogaster appears to be recently evolved. We, therefore, investigated possible functional consequences of sequence differences among these stripe 2 enhancers by P-element-mediated transformation. This analysis revealed that the eve stripe 2 enhancer from each of the four species drove reporter gene expression at the identical time and location in D. melanogaster embryos. Double staining of native eve protein and transgene mRNA in early embryos showed that the reporter gene mimicked native eve expression and, in every case, produced sharply defined stripes at the blastoderm stage that were coincident with eve stripe 2 protein. We argue that stripe 2 eve expression in Drosophila evolution can be viewed as being under constant stabilizing selection with respect to the location of the anterior and posterior borders of the stripe. We further hypothesize that the stripe 2 enhancer is functionally robust, so that its evolution may be governed by the fixation of both slightly deleterious and adaptive mutations in regulatory protein binding sites as well as in the spacing between binding sites. This view allows for a slow but continual turnover of functionally important changes in the stripe 2 enhancer.

Animals↗
PubMed1986

Cloning and identification of the gene coding for the 140-kd subunit of Drosophila RNA polymerase II.

Genomic clones of Drosophila melanogaster were isolated from a lambda library by cross-hybridization with the yeast gene coding for the 150-kd subunit of RNA polymerase II. Clones containing a region of approximately 2.0 kb with strong homology to the yeast gene were shown to code for a 3.9-kb poly(A)-RNA. Part of the coding region was cloned into an expression vector. A fusion protein was obtained which reacted with an antibody directed against RNA polymerase II of Drosophila. Peptide mapping of the fusion protein yielded a number of spots identical with spots derived from the 140-kd subunit of Drosophila RNA polymerase II. Sequence comparison of a segment of the Drosophila and the corresponding yeast clone yielded a high degree of homology at the protein level also, suggesting that we had isolated the gene coding for the 140-kd subunit of RNA polymerase II from Drosophila. In situ hybridization localized the DmRP140 gene at 88 A/B on chromosome 3 while the DmRP215 gene has previously been localized at 10 C on the X chromosome. Analysis of the transcripts (7.0 and 3.9 kb) in female and male flies shows dosage compensation for the transcription of the DmRP215 gene.

Journal Article↗
PubMed2000

Neurophysiological and behavioural evidence for an olfactory function for the dorsal organ and a gustatory one for the terminal organ in Drosophila melanogaster larvae.

Multicellular electrophysiological responses from the dorsal organ on the cephalic lobes of third instar Drosophila melanogaster larvae (wild-type Canton S) stimulated with a cold-trapped banana volatile extract showed that this structure has an olfactory function in the fruit fly. Responses of the dorsal organ were also recorded to constituents of the banana volatile extract as they eluted from a gas chromatographic column (GC-coupled dorsal organ electrophysiology). The active chemostimulants were identified as 2-heptanone, isoamyl alcohol, hexyl acetate, hexanol and hexyl butyrate by gas chromatography-coupled mass spectrometry. Applying the same recording system to the terminal organ sensilla, no responses were obtained to either the banana volatile bouquet or its constituents. By contrast, high frequency multicellular responses were recorded in response to touching the terminal organ with the gustatory stimuli KCl and grapefruit juice; responses were absent on similar stimulation of the dorsal organ with either NaCl or KCl. This suggests a role for olfaction by the dorsal organ and for gustation by the terminal organ in Drosophila larvae.In a 7-mm high wind tunnel with a thin 1.2% agar floor, the Drosophila larvae showed odour-conditioned upwind responses in an air stream of 0.1 m/s bearing banana volatiles. Drosophila larvae responded best to the odour of cut bananas. A 1:1 mixture of the banana odour constituents 2-heptanone and hexanol (at either 50 or 100 &mgr;g source dose each) proved as attractive as the known larval attractants propionic acid and isoamyl acetate on their own at 100 &mgr;g, whereas hexanol and 2-heptanone on their own at a 100 &mgr;g source dose were less attractive. The stronger behavioural response to the banana volatile bouquet and to the binary mixture serves to underline the multireceptor nature of the dorsal organ response to food odour in Drosophila.

Journal Article↗
PubMed2001

Drosophila as a model to study human brain degenerative diseases.

Drosophila has been an ideal system in which to identify molecules and define pathways involved in development, in part because of the powerful genetic approaches that are possible. Many of the molecules and pathways important in development in Drosophila are evolutionarily conserved between fly and human. With its highly evolved nervous system, amenability to genetic analysis, and the full genomic sequence available, Drosophila is a valuable tool for investigating and understanding the molecular mechanisms of neurodegenerative diseases.In order to have neurodegenerative Drosophila mutants, I screened EMS treated X chromosomes and P-element inserted 2nd and 3rd chromosomes in Drosophila for reduced life span and neurodegeneration. Twenty-one neurodegenerative mutants including bubblegum, spongecake, and eggroll were isolated and were named by virtue of their brain lesions. Each mutant has distinct pattern of degeneration in specific regions of the brain. Degeneration occurs in lamina and retina region in bubblegum. In spongecake vacuolization can only be seen in the optic lobe, especially in the medulla region. Multilamellated inclusions are wide-spread in the brain of eggroll. It showed not only do the pathologies iin fly brains resemble that of human diseases including Creutzfeldt Jakob disease, Tach-Sachs and Niemann-Pick disease, but the gene involved in the pathological pathway in the bubblegum mutant also functions as in the human adrenoleukodystrophy.

Journal Article↗
PubMed2026

Retention of a single Cenp-C gene in different syntenic locations in the montium group of Drosophila species.

Chromosome segregation in eukaryotes requires the orchestrated interaction of chromosomes with microtubules, mediated by the kinetochore multiprotein complex that assembles on chromosomal regions known as centromeres. In most eukaryotes, CenH3 and Cenp-C centromeric proteins are essential for centromere function. In Drosophila, the localization of CenH3 (or Cid in Drosophila) depends on its chaperone CAL1 and Cenp-C. Previous studies have shown that both Cid and Cenp-C underwent a coincident gene duplication and likely functional specialization in the Drosophila subgenus. Independently, Cid duplications led to three paralogs in the montium group (Sophophora subgenus). Here, we investigated whether this group also underwent parallel Cenp-C duplications by analyzing sequenced genomes of 23 montium group species. We identified Cenp-C genes in five distinct syntenic loci. Despite their distinct synteny, all but two montium group species (except D. birchii and D. vulcana) encode a single Cenp-C, whose phylogeny mirrors the species phylogeny, and all encode protein motifs indicative of intact Cenp-C function. These Cenp-C genes resulted from gene translocations or duplication followed by loss of the ancestral copy. Therefore, the co-retention of three Cid paralogs in the montium group did not result in coincident Cenp-C paralog co-retention. Analysis of the selective constraints in Cenp-C reveals more prominent positive selection in the Drosophila subgenus (with two retained Cenp-C paralogs) than in the Sophophora subgenus, including the montium group. Our work highlights differences in functional retention and potential specialization of CenH3 and Cenp-C, two of the most conserved eukaryotic centromeric proteins in Drosophila.

Journal Article↗
PubMed2005

REL1, a homologue of Drosophila dorsal, regulates toll antifungal immune pathway in the female mosquito Aedes aegypti.

Signaling by Drosophila Toll pathway activates two Rel/NF-kappaB transcription factors, Dorsal (Dl) and Dorsal-related immune factor (Dif). Dl plays a central role in the establishment of dorsoventral polarity during early embryogenesis, whereas Dif mediates the Toll receptor-dependent antifungal immune response in adult Drosophila. The absence of a Dif ortholog in mosquito genomes suggests that Dl may play its functional role in the mosquito Toll-mediated innate immune responses. We have cloned and molecularly characterized the gene homologous to Drosophila Dl and to Anopheles gambiae REL1 (Gambif1) from the yellow fever mosquito Aedes aegypti, named AaREL1. AaREL1 alternative transcripts encode two isoforms, AaREL1-A and AaREL1-B. Both transcripts are enriched during embryogenesis and are inducible by septic injury in larval and female mosquitoes. AaREL1 and AaREL2 (Aedes Relish) selectively bind to different kappaB motifs from insect immune gene promoters. Ectopic expression of AaREL1-A in both Drosophila mbn-2 cells and transgenic flies specifically activates Drosomycin and results in increased resistance against the fungus Beauveria bassiana. AaREL1-B acted cooperatively with AaREL1-A to enhance the immune gene activation in Aag-2 cells. The RNA interference knock-outs revealed that AaREL1 affected the expression of Aedes homologue of Drosophila Serpin-27A and mediated specific antifungal immune response against B. bassiana. These results indicate that the homologue of Dl, but not that of Dif, is a key regulator of the Toll antifungal immune pathway in A. aegypti female mosquitoes.

Aedes↗