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  • Drosophila melanogaster
  • Springer  (163)
  • Periodicals Archive Online (PAO)
  • 1995-1999  (96)
  • 1975-1979  (67)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Entomologia experimentalis et applicata 92 (1999), S. 289-294 
    ISSN: 1570-7458
    Keywords: parasitoid size ; Asobara ; host sex ; sex allocation ; Drosophila melanogaster ; parasitic castration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Parasitoid females are known to preferentially allocate female eggs to hosts with the higher resource value, usually leading to oviposition of female eggs in larger hosts and male eggs in smaller hosts. For koinobiont parasitoids, if male and female hosts are of equal size at time of oviposition, but differ in size in later developmental stages, the sex of the host could be used to indicate future resource value. Using parasitoids of the braconid genus Asobara, which are larval parasitoids of Drosophila, it is shown that parasitoids emerging from female hosts are larger than those from male hosts. Given this difference in resource value, ovipositing females should preferentially allocate female eggs to female hosts. An alternative strategy would be to decrease the difference in resource value between male and female hosts by castrating male hosts. The primary sex ratio of A. tabida in their two main host species does not differ between male and female hosts. In contrast to A. tabida, A. citri is known to partially castrate male hosts, but this does not decrease the size difference between male and female hosts. As in A. tabida, there is no difference in sex allocation to male and female hosts in A. citri. Despite the clear difference between the resource value of male and female hosts, these parasitoid species do not seem to make optimal use of this difference. They may not be able to discriminate between host sexes or, alternatively, there is a presently unknown fitness disadvantage to ovipositing in female hosts.
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  • 2
    ISSN: 1572-8889
    Keywords: Drosophila melanogaster ; larvae ; vision ; mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Foraging-stage third-instar larvae from most wild-type (normal) Drosophila melanogaster stocks are generally repelled by light. To identify factors that affect the larval photoresponse, we elucidated the effects of age, temperature, and time on the photoresponse of larvae from a wild-type Canton-S stock. In addition, we analyzed the larvae from the LI2 isofemale line, which are unresponsive to light in a photoassay. To determine whether LI2 larvae behave abnormally on other behavioral paradigms, in comparison to Canton-S controls, we tested larvae in taste and olfactory assays and observed them to determine whether they dispersed in a food source. Like Canton-S larvae, LI2 larvae and other isofemale lines whose progenitors were collected from the same natural population are responsive to taste and olfactory stimuli. Moreover, LI2 larvae disperse in the food source, as do Canton-S larvae tested in the dark. Larvae expressing parasbl mutations, which respond normally to light but not to chemical stimuli, do not disperse normally in the food source, suggesting that dispersal may be mediated by perception of chemical cues.
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  • 3
    ISSN: 1572-8889
    Keywords: Drosophila melanogaster ; Drosophila willistoni ; sexual behavior ; species isolation ; P elements
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Molecular analysis suggests that the pomace fly Drosophila melanogaster acquired the P family of transposable elements from another Drosophila species, D. willistoni. Since the two species are distantly related, it has been assumed that transmission of P element DNA from D. willistoni to D. melanogaster was mediated by a vector. The possibility of an alternative mode of transmission was assessed by characterizing the sexual behaviors of D. willistoni males and females, then observing D. willistoni and D. melanogaster males and females to see whether males from one species interacted sexually with females from the other species in a laboratory setting. We observed that D. melanogaster males court D. willistoni females vigorously and, in some cases, stimulate the females to be receptive to copulation. However, D. willistoni males perform relatively little courtship in response to D. melanogaster females and do not attempt to copulate. Thus, it is unlikely that sexual interactions effected the transmission of P element DNA from D. willistoni to D. melanogaster in the flies' natural habitat.
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  • 4
    Electronic Resource
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    Springer
    Journal of insect behavior 11 (1998), S. 47-57 
    ISSN: 1572-8889
    Keywords: Drosophila melanogaster ; D. hydei ; pupation height ; pH ; acidity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This paper describes the effects of the pH of the larval resource on the pupation height of two cosmopolitan species of Drosophila: D. melanogaster and D. hydei. The pH levels of “artificial media” were modified using solutions of acids (hydrochloric, acetic, and citric) and a positive relationship was found between pupation height and resource pH; i. e., the more acidic the resource, the closer the larvae pupated to the resource surface. The clearest trends were between pupation height and the final pH of the resource, the larvae apparently responding to the pH they encountered just before pupation commenced. When using natural resources a difference in pupation height was found between the types of fruits used but there was no obvious trend linking pupation height with pH. This suggests that some other environmental factor(s) was (were) overriding the effects of pH, and consequently resource pH may play only a minor role in determining pupation behavior in nature.
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  • 5
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    Springer
    Journal of insect behavior 8 (1995), S. 835-845 
    ISSN: 1572-8889
    Keywords: aging ; Drosophila melanogaster ; phototaxis ; locomotor activity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phototaxis was measured in young, middle-aged, and oldDrosophila melanogaster flies of both sexes. The apparatus allowed us to measure the tendency to go toward light, independently of the time needed to do so; under such conditions, phototaxis is dissociated from locomotor activity. The percentage of photopositive flies decreased slightly with age (93.96, 80.17, and 78.97%, respectively, in young, middle-aged, and old flies). Results are discussed in connection with previous data for which the tendency to go toward light and the time to do so were not dissociated.
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  • 6
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    Cellular and molecular life sciences 51 (1995), S. 73-76 
    ISSN: 1420-9071
    Keywords: Carcinogens ; genotoxicity testing ; Drosophila melanogaster ; somatic mutation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract To provide further background data on the wing spot somatic mutation and recombination assay, 10 selected carcinogens (acetamide, acrylamide, benzo(a)pyrene, cyclophosphamide, diethylstilbestrol, 4-nitroquinoline N-oxide, propyleneimine, safrole, thiourea, and o-toluidine) were tested in this assay. 72-h-old third-instar larvae, trans-heterozygous for 2 recessive wing cell markers:multiple wing hairs (mwh) andflare 3 (flr 3) were fed with 3 concentrations of each carcinogen during the rest of their development until pupation, and the genotoxic effects were measured as significant increases in the appearance of visible mutant hair clones on the adult wing blade. Our results show that 6 of the carcinogens tested produce significant increases in wing spot frequency, at least at one of the concentrations assayed. Benzo(a)pyrene, diethylstilbestrol, safrole and thiourea were the compounds that did not increase the incidence of mutant clones.
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  • 7
    ISSN: 1420-9071
    Keywords: Genotoxicity ; somatic cells ; spot size ; Drosophila melanogaster ; methyl methanesulfonate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The relationship between the induction of mutant clones and the time of mutagen treatment was studied in the somatic mutation and recombination test (SMART) in wing cells ofDrosophila melanogaster. Larvae trans-heterozygous for the recessive marker mutations multiple wing hairs (mwh) and flare (flr) were produced. Batches of these larvae were then treated with mutagen at different ages spanning all three larval instars. Methyl methanesulfonate was fed acutely for 2 h by immersing the larvae in a solution of the mutagen mixed with powdered cellulose. Wings of the surviving adult flies were mounted and scored for the presence of spots. The frequency and size of single and twin spots were recorded separately. Twin spots are produced exclusively by mitotic recombination, whereas single spots can results from various types of mutational and exchange events. There exists a clear correlation between time of induction and frequency as well as size of the single spots. In young larvae only few but very large spots are induced, whereas in older larvae the frequencies are considerably increased but the sizes are smaller. The twin spots show a different relationship. Practically no twin spots are found in very young and in very old larvae. The results demonstrate that in the wing spot test the optimal age of the larvae for mutagen treatment is 72 h.
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  • 8
    ISSN: 1432-1432
    Keywords: Esterase ; Gene cluster ; Pseudogene ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The α-esterase cluster ofD. melanogaster contains 11 esterase genes dispersed over 60 kb. Embedded in the cluster are two unrelated open reading frames that have sequence similarity with genes encoding ubiquitin-conjugating enzyme and tropomyosin. The esterase amino acid sequences show 37–66% identity with one another and all but one have all the motifs characteristic of functional members of the carboxyl/cholinesterase multigene family. The exception has several frameshift mutations and appears to be a pseudogene. Patterns of amino acid differences among cluster members in relation to generic models of carboxyl/cholinesterase protein structure are broadly similar to those among other carboxyl/cholinesterases sequenced to date. However the α-esterases differ from most other members of the family in: their lack of a signal peptide; the lack of conservation in cysteines involved in disulfide bridges; and in four indels, two of which occur in or adjacent to regions that align with proposed substrate-binding sites of other carboxyl/cholinesterases. Phylogenetic analyses clearly identify three simple gene duplication events within the cluster. The most recent event involved the pseudogene which is located in an intron of another esterase gene. However, relative rate tests suggest that the pseudogene remained functional after the duplication event and has become inactive relatively recently. The distribution of indels also suggests a deeper node in the gene phylogeny that separates six genes at the two ends of the cluster from a block of five in the middle.
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  • 9
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    Journal of molecular evolution 41 (1995), S. 430-439 
    ISSN: 1432-1432
    Keywords: Cytochrome proteins ; Molecular evolution ; Drosophila melanogaster ; Drosophila virilis ; Gene structure and sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cytochrome proteins perform a broad spectrum of biological functions ranging from oxidative metabolism to electron transport and are thus essential to all organisms. The b-type cytochrome proteins bind heme noncovalently, are expressed in many different forms and are localized to various cellular compartments. We report the characterization of the cytochrome b5 (Cyt-b) gene of Drosophila virilis and compare its structure to the Cyt-b gene of Drosophila melanogaster. As in D. melanogaster, the D. virilis gene is nuclear encoded and single copy. Although the intron/exon structures of these homologues differ, the Cyt-b proteins of D. melanogaster and D. virilis are approximately 75% identical and share the same size coding regions (1,242 nucleotides) and protein products (414 amino acids). The Drosophila Cyt-b proteins show sequence similarity to other b-type cytochromes, especially in the N-terminal heme-binding domain, and may be targeted to the mitochondrial membrane. The greatest levels of similarity are observed in areas of potential importance for protein structure and function. The exon sequences of the D. virilis Cyt-b gene differ by a total of 292 base changes. However, 62% of these changes are silent. The high degree of conservation between species separated by 60 million years of evolution in both the DNA and amino acid sequences suggests this nuclear cytochrome b5 locus encodes an essential product of the Drosophila system.
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  • 10
    ISSN: 1432-1432
    Keywords: Drosophila melanogaster ; Transposable element ; Mobilization ; Genomic stress ; Virus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To analyze the behavior of endogenous transposable elements under genomic stress, aDrosophila melanogaster inbred line was submitted to three kinds of viral perturbations. First, a retroviral plasmid containing the avian Rous Associated Virus type 2 (RAV-2) previously deleted for the viral envelope coding gene (env) was introduced by P element transformation into theDrosophila genome. An insertion of this avian retroviral sequence was detected byin situ hybridization in site 53C on polytene chromosome arm 2R. Second,Drosophila embryos were injected with RAV-2 particles produced by cell culture after transfection with the retroviral plasmid. Third, theDrosophila melanogaster inbred line was stably infected by the sigma native virus. It appears that neither the offspring of the flies in which the viral DNA was found integrated nor those from the infected sigma flies showed copia or mdgl element mobilization. Injection of the avian RAV-2 particles led, however, to the observation of somatic transpositions of mdgl element on the 2L chromosome, the copia element insertion pattern remaining stable. Thus, endogenous transposable elements show more instability in sublines injected with exogenous viral particles than in a transgenic subline containing a foreign viral insert, all transposable elements not being equally sensitive to such genomic stress.
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  • 11
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    Journal of molecular evolution 41 (1995), S. 155-160 
    ISSN: 1432-1432
    Keywords: TGF-β superfamily ; decapentaplegic gene ; Drosophila melanogaster ; Schistocerca americana ; Gene comparison
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Intercellular signaling molecules of the transforming growth factor-β (TGF-β) superfamily are required for pattern formation in many multicellular organisms. The decapentaplegic (dpp) gene of Drosophila melanogaster has several developmental roles. To improve our understanding of the evolutionary diversification of this large family we identified dpp in the grasshopper Schistocerca americana. S. americana diverged from D. melanogaster approximately 350 million years ago, utilizes a distinct developmental program, and has a 60-fold-larger genome than D. melanogaster. Our analyses indicate a single dpp locus in D. melanogaster and S. americana, suggesting that dpp copy number does not correlate with increasing genome size. Another TGF-β superfamily member, the D. melanogaster gene 60A, is also present in only one copy in each species. Comparison of homologous sequences from D. melanogaster, S. americana, and H. sapiens, representing roughly 900 million years of evolutionary distance, reveals significant constraint on sequence divergence for both dpp and 60A. In the signaling portion of the dpp protein, the amino acid identity between these species exceeds 74%. Our results for the TGF-β superfamily are consistent with current hypotheses describing gene duplication and diversification as a frequent response to high levels of selective pressure on individual family members.
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  • 12
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    Development genes and evolution 182 (1977), S. 69-74 
    ISSN: 1432-041X
    Keywords: Drosophila melanogaster ; Male foreleg disk ; Capacity of transdetermination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In the male foreleg disk ofDrosophila melanogaster the cells capable of transdetermination are clustered in a specific region within the upper half of the disk. Cells outside this region cannot transdetermine under any of the experimental conditions thus far applied. Transdetermination occurs when cells capable of transdetermination are stimulated to a certain extent of additional proliferation. This can be achieved either by exposing these cells at a wound surface of an intact fragment, or by dissociation.
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  • 13
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    Development genes and evolution 187 (1979), S. 151-165 
    ISSN: 1432-041X
    Keywords: Oogenesis ; Embryogenesis ; Two-dimensional gels ; Protein synthesis ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Protein synthesis in egg follicles and blastoderm embryos ofDrosophila melanogaster has been studied by means of two-dimensional gel electrophoresis. Up to 400 polypeptide spots have been resolved on autoradiographs. Stage 10 follicles (for stages see King, 1970) were labelled in vitro for 10 to 60 min with35S-methionine and cut with tungsten needles into an anterior fragment containing the nurse cells and a posterior fragment containing the oocyte and follicle cells. The nurse cells were found to synthesize a complex pattern of proteins. At least two proteins were detected only in nurse cells but not in the oocyte even after a one hour labelling period. Nurse cells isolated from stages 9, 10 and 12 follicles were shown to synthesize stage specific patterns of proteins. Several proteins are synthesized in posterior fragments of stage 10 follicles but not in anterior fragments. These proteins are only found in follicle cells. No oocyte specific proteins have been detected. Striking differences between the protein patterns of anterior and posterior fragments persist until the nurse cells degenerate. In mature stage 14 follicles, labelled in vivo, no significant differences in the protein patterns of isolated anterior and posterior fragments could be detected; this may be due to technical limitations. At the blastoderm stage localized synthesis of specific proteins becomes detectable again. When blastoderm embryos, labelled in vivo, are cut with tungsten needles and the cells are isolated from anterior and posterior halves, differences become apparent. The pole cells located at the posterior pole are highly active in protein synthesis and contribute several specific proteins which are found exclusively in the posterior region of the embryo. In this study synthesis of specific proteins could only be demonstrated at those developmental stages which are characterized by the presence of different cell types within the egg chamber, while no differences were detected when stage 14 follicles were cut and anterior and posterior fragments analyzed separately. The differences in the pattern of protein synthesis by pole cells and blastoderm cells indicate that even the earliest stages of determination are reflected by marked changes at the biochemical level.
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  • 14
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    Cellular and molecular life sciences 51 (1995), S. 744-748 
    ISSN: 1420-9071
    Keywords: Morphometry ; geographic races ; chromosome transfers ; Drosophila melanogaster ; natural populations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The genetic basis of three morphological traits (ovariole number, sternopleural bristle number and wing length) ofDrosophila melanogaster has been investigated in natural populations that show great differences in these traits, i. e. Bordeaux (France) and Loua (Congo). F1 and F2 crosses, and chromosome substitutions between these two populations, were analysed. Maternal and/or X chromosome effects were found for sternopleural bristle number and wing length. For all traits, significant effects from each of the three chromosomes were found, but in general only one or two chromosomes had a major effect. Moreover, in all cases significant interactions between chromosomes were observed, suggesting the existence of epistatic effects. Our results are discussed and compared to those obtained from the analysis of selected laboratory strains.
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  • 15
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    Development genes and evolution 206 (1996), S. 277-280 
    ISSN: 1432-041X
    Keywords: Key words Nervous system development ; Metamorphosis ; Phagocytosis ; Glial cells ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Using electron microscopy we demonstrate that degenerating neurons and cellular debris resulting from neuronal reorganization are phagocytosed by glial cells in the brain and nerve cord of the fruitfly Drosophila melanogaster during the first few hours following pupariation. At this stage several classes of glial cells appear to be engaged in intense phagocytosis. In the cell body rind, neuronal cell bodies are engulfed and phagocytosed by the same glial cells that enwrap healthy neurons in this region. In the neuropil, cellular debris in tracts and synaptic centres resulting from metamorphic re-differentiation of larval neurons is phagocytosed by neuropil-associated glial cells. Phagocytic glial cells are hypertrophied, produce large amounts of lysosome-like bodies and contain a large number of mitochondria, condensed chromatin bodies, membranes and other remains from neuronal degeneration in phagosomes.
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  • 16
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    Development genes and evolution 208 (1998), S. 106-112 
    ISSN: 1432-041X
    Keywords: Key words Apoptosis ; Oogenesis ; Nurse cells ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  In Drosophila a remarkable feature of oogenesis is the regression of the nurse cells after dumping their cytoplasmic contents into the oocyte. We have studied the nature of this process at the late stages of egg chamber development. In egg chambers DAPI staining shows highly condensed chromatin from stage 12 and TUNEL labelling shows DNA fragmentation up to stage 14. Gel electrophoresis of the end-labelled DNA, extracted from isolated egg chambers at the same stages of development, shows a ladder typical of apoptotic nuclei. This provides evidence that, during Drosophila oogenesis, the nurse cells undergo apoptosis. Apoptotic nuclei have also been detected in dumping-defective egg chambers, indicating that the cytoplasmic depletion of nurse cells is concurrent with but apparently not the cause of the process.
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  • 17
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    Development genes and evolution 183 (1977), S. 249-268 
    ISSN: 1432-041X
    Keywords: Pattern-formation ; Embryogenesis ; Maternal-effect mutants ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The mutationbicaudal (Bull, 1966) causes embryos to develop a longitudinal mirror image duplication of the posteriormost abdominal segments, while head and thorax are missing. These embryos occur with varying frequencies among eggs laid by mutant females, irrespective of the paternal genotype. Recombination and deletion mapping indicate thatbicaudal (bic) is a recessive, hypomorphic, maternal-effect mutation mapping at a single locus on the second chromosome ofDrosophila melanogaster close tovg (67.0±0.1). The frequency of bicaudal embryos depends on the age of the mother, her genetic constitution and the temperature at which she is raised. Best producers are very young females hemizygous forbic (bic/Df(2)vg B ) at 28° C. Under these conditions 80% to 90% of the eggs which differentiate can show the bicaudal embryo phenotype. Upon ageing of the mother the frequency of bicaudal embryos declines rapidly, and most of the eggs develop the normal body pattern. Temperature shift experiments suggest a temperature-sensitive period at the onset of vitellogenesis. The mutation causes several types of abnormalities in the segment pattern of theDrosophila embryo, which are interpreted as various degrees of expression of the mutant character. The most frequent abnormal phenotype is the symmetrical bicaudal embryo with one to five abdominal segments duplicated. Less frequent are asymmetrical types, in which the smaller number of segments is always in the anterior reversed part. Other phenotypes are embryos with missing or rudimentary heads, and embryos with irregular gaps in the segment pattern. In bicaudal embryos, the pole cells, formed at the posterior pole of the egg prior to blastoderm formation, are not duplicated at the anterior. The significance of thebicaudal phenotypes for embryonic pattern-formation inDrosophila is discussed.
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  • 18
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    Development genes and evolution 183 (1977), S. 165-169 
    ISSN: 1432-041X
    Keywords: Clones ; Nervous system ; Shibire ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mitotic recombination was induced, by X-irradiation at the blastoderm stage, in flies heterozygous for one of the temperature-sensitive paralytic mutationsshibire andtp-2. The results show that these mutations can be used to detect the presence of clones in the central nervous system through the temperature-sensitive paralysis of individual legs. Mitotic recombination can also be used to examine the effects of these mutations in the peripheral nervous system; shibire is thus shown to affect the function of sensory neurons.
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  • 19
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    Development genes and evolution 180 (1976), S. 107-119 
    ISSN: 1432-041X
    Keywords: Drosophila melanogaster ; Cell lines ; Isoenzymes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Our previous isoenzyme investigation ofDrosophila melanogaster cell lines in vitro has been completed with twelve further enzyme systems. The “enzyme profiles” seem to be in good agreement with a previous hypothesis concerning the precise origin of these cell lines (probably from imaginal discs or nervous tissues). Our results have been summarized with reference to the biochemical genetic map ofDrosophila melanogaster in order to consider a possible functional organization of the genome.
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  • 20
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    Development genes and evolution 184 (1978), S. 41-56 
    ISSN: 1432-041X
    Keywords: Drosophila melanogaster ; Female germ line ; Mosaics ; Stem cell divisions ; Metafemale ; Sterility
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Our report presents an analysis of the development and dynamics of the female germ line inDrosophila. Females were produced that were mosaic either for attached-X chromosomes $$(\widehat{XX})$$ and a ring-X (triplo-X-diplo-X), or for $$\widehat{XX}$$ and a marked Y-chromosome $$(\widehat{XX}/Y - \widehat{XX}/O)$$ . The germ-line and genitalia of these females were analysed by direct microscopic observation or by examination of the progeny. Eggs derived from triplo-X germ cells were hardly capable of supporting development, with most of the zygotes dying during embryonic development. The analysis of the germ line was therefore carried out mainly by direct observation of histochemically stained developing oocytes in the ovaries of mosaic females. The total germ cell population of both ovaries of a female was mosaic in 22–29% of the tested animals. From this frequency of mosaicism we estimated the number of functional primordial germ cells to be betwen 3 and 6 cells at the blastoderm stage. At this stage the cell lineages for the left and right ovary are not yet separated. The germ cell population of individual ovarioles was frequently mosaic which shows that the few stem cells in an ovariole are recruited as a group and are not clonal descendants of a single ancestor cell per ovariole. An analysis of the sequential pattern of oocyte-nurse cell cysts in mosaic ovarioles revealed that neighbouring cysts tend to be of the same genotype. This suggests that the stem cells of the adult ovaries preferentially divide in bursts, one of them giving rise to two, three and sometimes even more cystocytes in a row. In addition, the foci for lethality and sterility of the triplo-X condition were determined. Non-mosaic triplo-X females (metafemales) are hardly viable and invariably sterile. Using our mosaics, the focus forlethality could be mapped to a region very near the ventral prothoracic discs. The focus forsterility resides in the genitalia, since flies with triplo-X genitalia never laid any eggs, regardless of the genotype of their ovaries.
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  • 21
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    Development genes and evolution 187 (1979), S. 167-177 
    ISSN: 1432-041X
    Keywords: Pyrimidine biosynthesis ; rudimentary mutants ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The X-linkedrudimentary (r) mutants ofDrosophila melanogaster are pyrimidine auxotrophs and require exogenous pyrimidines (Nørby, 1970; Falk, 1976). We have established a set ofrudimentary cell lines that are derived from embryos, homozygous for eitherr 1 orr 36. The enzymatic activities of the pyrimidine synthesizing enzymes were measured in the mutant lines. We have further investigated the nutritional requirements of the mutant cells in vitro by using a pyrimidine free culture medium. Ther 1 cell lines were found to express 3–7%dihydroorotase (DHOase) activity as compared to a wildtype cell line. Reducedaspartate transcarbamylase (ATCase) activity was measured in somer 1 cell lines whereas wildtypecarbamylphosphate synthetase (CPSase) activity is expressed in allr 1 cell lines. Ther 36 cell line expresses wildtype activity ofDHOase andCPSase. ATCase activity was found to be reduced to 10% of the wildtype activity. The mutant cell lines do not proliferate in pyrimidine free minimal medium and cell proliferation is obtained by the addition of crude RNA. Proliferation of ther 1 cells is restored by the supplementation of the minimal medium withdihydroorotate whereas proliferation of ther 36 cells is restored by supplementation with eitherdihydroorotate orcarbamylaspartate. The results demonstrate that therudimentary phenotypesr 1 andr 36 are expressed at the cellular level and that the two mutant cell types behave as cellular pyrimidine auxotrophs in vitro.
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  • 22
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    Development genes and evolution 208 (1998), S. 578-585 
    ISSN: 1432-041X
    Keywords: Key words Glial cells missing ; Drosophila melanogaster ; GAL4-UAS system ; Ectopic expression ; Cell fate determination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The glial cells missing (gcm) gene encodes an essential transcription factor that converts neuronal precursor cells to glial fate in the Drosophila nervous system. In this study, we tested effects of gcm ectopic expression on fate of non-neural cells. When gcm expression was continuously induced in epidermal cells from around stage 9, these cells started to exhibit mesenchymal cell morphology at stage 13, which was preceded by the onset of expression of Repo, a glial marker. The morphological change was coincident with loss of expression of an epidermal cell-adhesion molecule. In addition to the epidermis, fate of mesodermal cells was also affected by gcm ectopic expression. These findings suggest that gcm can convert gene expression and cell morphology even outside the neuroectoderm.
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  • 23
    ISSN: 1432-041X
    Keywords: multi sex combs ; Germline development ; Cell proliferation ; Polycomb group ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We present a genetic analysis showing that the Drosophila melanogaster gene multi sex combs (mxc; Santamaria and Randsholt 1995) is needed for proliferation of the germline. Fertility is the feature most easily affected by weak hypomorphic mutations of this very pleiotropic locus. Pole cell formation and early steps of gonadogenesis conform to the wild-type in embryos devoid of zygotic mxc + product. mxc mutant gonad phenotypes and homozygous mxc germline clones suggest a role for mxc + in control of germ cell proliferation during the larval stages. mxc + requirement is germ cell autonomous and specific in females, whilst in males mxc + product is also needed in somatic cells of the gonads. Although mxc can be classified among the Polycomb group (Pc-G) of genes, negative trans-regulators of the ANT-C and BX-C gene complexes, germline requirement for mxc appears independent of a need for other Pc-C gene products, and mxc gonad phenotypes are different from those induced by mutations in BX-C genes. We discuss the possible functions of the mxc + product which helps to maintain homeotic genes repressed and prevents premature larval haemocyte differentiation and neoplasic overgrowth, but promotes growth and differentiation of male and female gonads.
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  • 24
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    Development genes and evolution 180 (1976), S. 73-77 
    ISSN: 1432-041X
    Keywords: Ecdysones ; Imaginal discs ; Fat body ; Drosophila melanogaster
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    Topics: Biology
    Notes: Summary The effect of suboptimal levels of α-ecdysone on the differentiation in vitro ofDrosophila melanogaster wing discs was enhanced by the addition of larval fat body to the cultures. However, similar experiments with β-ecdysome showed no enhancement. It is suggested that a partial conversion of α-ecdysone to β-ecdysone by the fat body may well account for these results.
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  • 25
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    Development genes and evolution 181 (1977), S. 309-320 
    ISSN: 1432-041X
    Keywords: Drosophila melanogaster ; Male foreleg disc ; Pattern regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1. The developmental potentials of the four quadrants of the male foreleg disc ofDrosophila melanogaster were analysed by culturing excised quadrants for 3 days and 10 days in adult hosts prior to metamorphosis. 2. The cultured pieces underwent different types of pattern regulation in a circular direction. The upper medial piece was able to regenerate the missing structures of the disc, thus confirming the findings of earlier reports. The three remaining pieces could undergo pattern duplication in mirror-image symmetry. The lower medial piece revealed in addition a slight capacity for regeneration from the vertical cut surface. 3. The duplicating pieces differed markedly in their frequencies of pattern duplication: duplications occurred with very high frequencies in lower medial pieces, with intermediate frequencies in upper lateral pieces, and with very low frequencies in lower lateral pieces. 4. Both lower lateral and upper lateral pieces underwent a progressive loss of most markers with increasing culture time. 5. Claws were regenerated solely by upper medial pieces. 6. Transdetermined structures, too, were encountered only in upper medial pieces. 7. The results are discussed with respect to the two major current models of pattern regulation in imaginal discs, the “gradient model” and the “clock model”. 8. It is suggested that the differences in the frequencies of pattern duplication reflect the unequal spacing of circular positional values within the three duplicating quadrants. Under this assumption the data indicate a progressive decrease in the density of circular positional values with increasing distance from the upper medial quadrant of the disc.
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  • 26
    ISSN: 1432-1939
    Keywords: Key words Development ; Natural hyperthermia ; Heat shock proteins ; Hsp70 ; Drosophila melanogaster
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    Topics: Biology
    Notes: Abstract  We demonstrate that natural heat stress on wild larval Drosophila melanogaster results in severe developmental defects in 〉10% of eclosing adults, and that increased copy number of the gene encoding the major inducible heat shock protein of D. melanogaster, Hsp70, is sufficient to reduce the incidence of such abnormalities. Specifically, non-adult D. melanogaster inhabiting necrotic fruit experienced severe, often lethal heat stress in natural settings. Adult flies eclosing from wild larvae that had survived natural heat stress exhibited severe developmental anomalies of wing and abdominal morphology, which should dramatically affect fitness. The frequency of developmental abnormalities varied along two independent natural thermal gradients, exceeding 10% in adults eclosing from larvae developing in warm, sunlit fruit. When exposed to natural heat stress, D. melanogaster larvae with the wild-type number of hsp70 genes (n=10) developed abnormal wings significantly more frequently than a transgenic sister strain with 22 copies of the hsp70 gene.
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  • 27
    ISSN: 1573-4927
    Keywords: aldehyde oxidase ; xanthine dehydrogenase ; Drosophila melanogaster ; molybdenum
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Two new mutants, deficient in aldehyde oxidase and xanthine dehydrogenase, have been isolated from a wild-type stock of Drosophila melanogaster and have been provisionally termed lxd c and lxd d, respectively, as both mutants appear to be allelic with lxd (low xanthine dehydrogenase). An analysis has been made of the effects of dietary molybdenum on lxd, lxd c, lxdd, lao (low aldehyde oxidase), mal (maroon-like eye color), and pac (Pacific) wild-type flies. On the lower dietary levels of 10 −3 M and 10 −2 M molybdenum, increases in specific activity of both enzymes were observed only in lxd. Furthermore, two- to three-fold increases in specific activity of both enzymes occurred in all strains, except mal, when cultured on 5×10 −2 M molybdenum. The lxd and lxd c strains failed to survive on this high concentration of the ion. Similar concentrations of molybdenum had no effect in vitro. An extra electrophoretic band of xanthine dehydrogenase was observed on polyacrylamide gel from extracts of wild-type flies cultured on certain levels of molybdenum, but its appearance was not always correlated with the increases in specific activity.
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  • 28
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    Biochemical genetics 14 (1976), S. 357-371 
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; poly(A)-containing RNA
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The size range of poly(A)-containing RNA from Drosophila melanogaster embryos has been estimated by hybridization with 3H-labeled poly(U) and subsequent fractionation on sucrose gradients. The median size of nuclear poly(A)-containing RNA is about 30 S (6000 nucleotides), and the median size of cytoplasmic poly(A)-containing RNA is about 17 S (1800 nucleotides). The relationship of these sizes to messenger RNA needed to code for protein and to the length of DNA contained in a chromomere is discussed.
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  • 29
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    Biochemical genetics 14 (1976), S. 259-270 
    ISSN: 1573-4927
    Keywords: GTP cyclohydrolase ; Drosophila melanogaster ; pteridines ; dihydroneopterin triphosphate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The first enzyme (named GTP cyclohydrolase) in the pathway for the biosynthesis of pteridines has been partially purified from extracts of late pupae and young adults of Drosophila melanogaster. This enzyme catalyzes the hydrolytic removal from GTP of carbon 8 as formate and the synthesis of 2-amino-4-hydroxy-6-(d-erythro-1′,2′,3′-trihydroxypropyl)-7,8-dihydropteridine triphosphate (dihydroneopterin triphosphate). Some of the properties of the enzyme are as follows: it functions optimally at pH 7.8 and at 42 C; activity is unaffected by KCl and NaCl, but divalent cations (Mg2+, Mn2+, Zn2+, and Ca2+) are inhibitory; the K m for GTP is 22 μm; and the molecular weight is estimated at 345,000 from gel filtration experiments. Of a number of nucleotides tested, only GDP and dGTP were used to any extent as substrate in place of GTP, and these respective compounds were used only 1.8% and 1.5% as well as GTP.
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  • 30
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    Biochemical genetics 14 (1976), S. 611-617 
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; phenol oxidases ; spectrophotometry ; electrophoresis ; suppression ; ribosomal proteins
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract An interaction between the lozenge gene and the suppressor of forked gene of Drosophila melanogaster has been investigated both spectrophotometrically and electrophoretically. The nature of this interaction is such that certain lozenge alleles appear to be phenotypically suppressed while others are enhanced or unaffected, and the results reported demonstrate that the effect can clearly be observed at the biochemical level. Earlier observations have suggested that the suppressor of forked gene codes for a ribosomal protein, and this hypothesis is discussed.
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  • 31
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    Biochemical genetics 15 (1977), S. 93-100 
    ISSN: 1573-4927
    Keywords: alcohol dehydrogenase (ADH) ; genetic polymorphism ; selection ; Drosophila melanogaster
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract In the natural populations +Tüb, +Prov, and +Rov, similar Adh F allele frequencies occur (q F=0.11, 0.18, and 0.08, respectively). However, there is a discrepancy in that the Adh F allele in +Tüb is closely linked to the lethal factor 1(2)Stm, which reduces relative fitness of the F phenotype to zero. In spite of this, polymorphism is maintained also in +Tüb, because the heterozygotes are superior to the homozygous S type (relative fitness=0.88). Under laboratory culture conditions, in +Tüb the relative fitness of the S genotype further decreases to 0.6. After outcrossing the lethal factor, relative fitnesses for S, FS, and F become 0.6, 1, and 0.48, respectively, implying that fitness for S remains the same. Relative values for S, FS, and F in +Prov, not affected by the lethal factor, are calculated by the maximum average fitness method to be 1, 1.2, and 0.2 under the assumption that heterozygous FS are similarly superior to S as in the natural +Tüb population and all allele frequencies found are stable equilibrium values.
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  • 32
    ISSN: 1573-4927
    Keywords: l-glycerol-3-phosphate dehydrogenase (α-GPDH) ; isozymes ; development ; Drosophila melanogaster
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The basis for the differentiation of l-glycerol-3-phosphate dehydrogenase (α-GPDH) into larval and adult isozymes in Drosophila melanogaster was investigated by the correlation of a lack of appearance of each isozyme during development within Drosophila bearing α-GPDH “null” alleles and by the study of a putative conversion factor. Conversion studies indicate the presence of a heat-labile RNase-resistant conversion factor present in crude larval extracts with the ability to convert GPDH-1 to GPDH-2 and GPDH-3 but not vice versa. In addition, “null” mutations at the Gpdh locus obliterate all isozymatic species of α-GPDH in all developmental stages. These observations suggest that all α-GPDH isozymes are the product of a single structural gene and that the multiple forms of this enzyme arise during successive developmental stages through an epigenetic modification of the primary Gpdh + polypeptide. Finally, observations are reported which bear on the functional divergence of the α-glycerophosphate cycle in the adult and larval stage of development.
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  • 33
    ISSN: 1573-4927
    Keywords: allozymes ; thermostability ; alcohol dehydrogenase ; Drosophila melanogaster ; natural populations
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Drosophila melanogaster collected from natural populations were examined fo thermostability variants within electrophoretic mobility classes of two enzymes. In alcohol dehydrogenase, two discrete forms of the “slow” allozyme and three discrete forms of the “fast” allozyme were revealed by postelectrophoretic treatments ranging from 15 sec at 40 C to 40 sec at 43 C. All variants have been mapped to within 0.7 unit of the Adh locus. Results of a geographic survey indicate that two alleles giving rise to fast-moderate and slow-moderate allozymes are common everywhere; other variants have a collective frequency ranging from 0% to 7%. In a test of the possibility that the rare Adh alleles could be generated by intragenic recombination between the two common alleles, electrophoresis and heat treatment of progeny recombinant for flanking markers of Adh revealed no new allozymes. Among 27 stocks containing slow α-glycerophosphate dehydrogenase allozymes and 109 fast stocks, heat treatments revealed no additional variation.
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  • 34
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; GTP cyclohydrolase ; development ; pteridine biosynthesis ; mutants
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The reaction catalyzed by GTP cyclohydrolase is the first unique step of pteridine biosynthesis in Drosophila melanogaster and is therefore likely to be an important control point. GTP cyclohydrolase activity varies during development, showing two distinct peaks of activity—one at pupariation and a much larger peak at emergence. Most of the early pupal enzyme is located in the body region, whereas in late pupal and early adult life most of the activity is found in the head. Mixing experiments indicate that developmental changes in activity are not due to changes in the level of a direct effector of GTP cyclohydrolase. The mutants raspberry and prune show an increased GTP cyclohydrolase activity at pupariation relative to wild type, but a decreased enzyme activity at emergence. The changes in GTP cyclohydrolase activity are reflected in changes in pteridine levels in these mutants. Several lines of evidence suggest that neither locus is the structural gene for GTP cyclohydrolase. The raspberry and prune gene products may play a specific role in regulating GTP cyclohydrolase activity during development.
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  • 35
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    Biochemical genetics 16 (1978), S. 1113-1134 
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; isozymes ; position effect ; segmental aneuploidy
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A deoxyribonuclease, called DNase-1, that is active at acid pH in the presence of EDTA has been studied in Drosophila melanogaster. The locus for the enzyme maps genetically to 61.8 on the right arm of the third chromosome. Cytogenetically, DNase-1 has been localized to within five to ten bands between 90C-2 and 90E. This analysis utilizes both electrophoretic variants and the Y-autosome translocations of Lindsley et al. (1972). DNase-1 is present in all stages of the life cycle, and the paternal genome actively contributes DNase-1 to the embryo between 0 and 1 hr after fertilization.
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  • 36
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    Biochemical genetics 16 (1978), S. 159-170 
    ISSN: 1573-4927
    Keywords: alcohol dehydrogenase ; allozyme properties and amounts ; Drosophila melanogaster
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Among strains of Drosophila melanogaster each derived from a single fertilized female taken from natural populations, there is variation in both alcohol dehydrogenase (ADH) activity and the amount of ADH protein. The correlation between ADH activity and number of molecules over all strains examined is 0.87 or 0.96 in late third instar larvae depending on whether the substrate is 2-propanol or ethanol. With respect to the two common electrophoretic allozymic forms, F and S, segregating in these populations, the FF strains on the whole have higher ADH activities and numbers of ADH molecules than the SS strains. Over all strains examined, enzyme extracts from FF strains have a mean catalytic efficiency per enzyme molecule higher than that of enzyme extracts from SS strains when ethanol is the substrate, and much higher when 2-propanol is the substrate. One FF strain had an ADH activity/ADH protein ratio characteristic of SS strains.
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  • 37
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; alcohol tolerance ; alcohol utilization ; alcohol dehydrogenase ; aldehyde oxidase ; allozymes
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Alcohol dehydrogenase is necessary for ethanol detoxification and metabolic utilization. It has been generally assumed that aldehyde oxidase (AO) produced by the Aldox locus (3–56.7) is necessary for a further transformation of acetaldehyde into acetate. We find that various mutant strains (ma-l or Aldox n) which do not produce an active enzyme show about the same tolerance to alcohol as do wild strains. This physiological paradox is probably to be explained by the discovery of another locus (not localized) which produced a small amount of AO in all tested strains. The adaptive significance of the genetically polymorphic Aldox locus is probably to be looked for in physiological pathways other than ethanol metabolism.
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  • 38
    ISSN: 1573-4927
    Keywords: nonelectrophoretic structural variability ; Drosophila melanogaster ; phosphoglucomutase ; genetic polymorphism ; heat denaturation study
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    Notes: Abstract A simple procedure is described to detect genetic heterogeneity within electrophoretic classes at a locus in Drosophila, based on electrophoresis and heat denaturation studies. Temperature-resistant (tr) and temperature-sensitive (ts) isoelectrophoretic alleles at the phosphoglucomutase locus (Pgm) are present at polymorphic frequencies in natural and in laboratory populations of Drosophila melanogaster.
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  • 39
    ISSN: 1573-4927
    Keywords: sepiapterin synthase ; variegation ; purple ; Drosophila melanogaster ; pteridine eye pigments ; drosopterin
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A variegated position effect on the autonomous gene, purple, has been studied enzymologically in Drosophila melanogaster. Sepiapterin synthase, the enzyme system associated with pr +, was examined for activity in different developmental stages of the fly. The results indicate that T(Y:2) pr c5, cn/prc4 cn flies (flies in which pr + has been translocated and which exhibit variegation) have a reduced amount of enzyme activity as compared with both Oregon-R and pr 1 flies. This reduction in activity was not found in larval stages, which suggests that the inactivation process probably occurs in late larval or early pupal stages. The phenotype of the variegated adult has white eyes with red-colored spots and patches where drosopterins occur. The phenotype of the fly carrying the translocation is modified by the presence of additional Y chromosomes. This extends the observation from other systems that extra heterochromatin acts to suppress the variegated position effect. The advantages of studying the variegation by measuring enzyme activity, as well as the phenotypic expression, are several; for example, the developmental time at which variegation occurs may be estimated even though drosopterin synthesis is not occurring.
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  • 40
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    Biochemical genetics 17 (1979), S. 1131-1144 
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; enzyme polymorphism ; G6PD ; 6PGD ; enzyme activity
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The electrophoretic variants of G6PD and 6PGD isolated from the Bogota Drosophila melanogaster population were characterized developmentally and biochemically. Changes in in vitro enzyme activity during development were comparable to those found for other dehydrogenases: an increase in the larval and adult stage and a decrease in the pupal stage. During the whole life cycle the “S” enzyme of both loci showed a higher activity than the “F” enzyme. MgCl2 had a stimulating effect on the activity of both enzymes whereas their heat stability was decreased. The allozymes of 6PGD had different Vmax's but were comparable with respect to Km values, pH optimum, and stability at 45 C. the allozymes of G6PD showed different Vmax's and differed in stability at 35 C, but had similar Km values and pH optima. As the difference in stability was probably due to differences in molecular structure of the allozymes, the differences in activity found at high pH and high MgCl2 concentration were most probably due to this difference in stability.
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  • 41
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    Molecular and cellular biochemistry 158 (1996), S. 149-159 
    ISSN: 1573-4919
    Keywords: tubulin ; C-terminal regulatory domain ; DMAP-85 ; Drosophila melanogaster ; microtubule affinity columns
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The interaction of microtubule associated proteins (MAPs) with the microtubule system has been characterized in depth in neuronal cells from various mammalian species. These proteins interact with well-defined domains within the acidic tubulin carboxyl-terminal regulatory region. However, there is little information on the mechanisms of MAPs-tubulin interactions in nonmammalian systems. Recently, a novel tau-like protein designated as DMAP-85 has been identified in Drosophila melanogaster, and the regulation of its interactions with cytoskeletal elements was analyzed throughout different developmental stages of this organism. In this report, the topographic domains involved in the binding of DMAP-85 with tubulin heterodimer were investigated. Affinity chromatography of DMAP-85 in matrixes of taxol-stabilized microtubules showed the reversible interaction of DMAP-85 with domains on the microtubular surface. Co-sedimentation studies using the subtilisin-treated tubulin (S-tubulin) indicated the lack of association of DMAP-85 to this tubulin moiety. Moreover, studies on affinity chromatography of the purified 4 kDa C-terminal tubulin peptide bound to an affinity column, confirmed that DMAP-85 interacts directly with this regulatory domain on tubulin subunits. Further studies on sequencial affinity chromatography using a calmodulin affinity column followed by the microtubule column confirmed the similarities in the interaction behavior of DMAP-85 with that of tau. DMAP-85 associated to both calmodulin and the microtubular polymer. These studies support the idea that the carboxyl-terminal region on tubulin constitutes a common binding domain for most microtubule-interacting proteins.
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  • 42
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    Biochemical genetics 13 (1975), S. 603-613 
    ISSN: 1573-4927
    Keywords: transport mutants ; eye color mutants ; kynurenine ; Drosophila melanogaster ; Malpighian tubules
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Kynurenine-H 3 transport and conversion to 3-hydroxykynurenine were studied in organ culture using the Malpighian tubules and developing eyes from wild type and the eye color mutants w, st, 1td, ca, and cn of Drosophila melanogaster. Malpighian tubules from wild type have the ability to concentrate kynurenine and convert it to 3-hydroxykynurenine. The tubules from w, st, 1td, and ca are deficient in the ability to transport kynurenine, as are the eyes of the mutants w, st, and 1td. This defect in kynurenine transport provides a physiological explanation for the phenotypic properties of the mutants. The relationship of these measurements to previous observations on these eye color mutants is discussed and the transport defect hypothesis is consistently supported. We have concluded that several of the eye color mutants in Drosophila are transport mutants.
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  • 43
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; GTP cyclohydrolase ; pteridine biosynthesis ; development ; mutants
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    Notes: Abstract The enzyme guanosine triphosphate cyclohydrolase (GTP cyclohydrolase), which in bacteria is known to be the first enzyme in the biosynthetic pathway for the synthesis of pteridines, has been discovered in extracts of Drosophila melanogaster. Most of the enzyme (80%) is located in the head of the adult fly. An analysis of enzyme activity during development in Drosophila has revealed the presence of a relatively small peak of activity at pupariation and a much larger peak that appears at about the time of eclosion. Enzyme activity declines rapidly as the fly ages. Analyses for the production of the typical pteridine pigments of Drosophila have indicated that the small peak of GTP cyclohydrolase activity evident at pupariation coincides with the appearance of isoxanthopterin, sepiapterin, and pterin, and the larger peak at eclosion roughly corresponds to the accumulation of drosopterin as well as to the appearance in larger amounts of pterin and sepiapterin. These observations strongly suggest that in Drosophila, like bacteria, GTP cyclohydrolase is involved in the biosynthesis of pteridines. Analyses of a variety of zeste mutants of Drosophila melanogaster have shown that these mutants all contain GTP cyclohydrolase equal approximately to the amount found in the wild-type fly. These observations do not support the suggestions made by Rasmusson et al. (1973) that zeste is the structural locus for GTP cyclohydrolase.
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  • 44
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    Biochemical genetics 16 (1978), S. 333-342 
    ISSN: 1573-4927
    Keywords: β-hydroxy acid dehydrogenase ; chromosome ; dosage compensation ; Drosophila melanogaster
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A mutant Had nl was induced in Drosophila melanogaster and found to be deficient in β-hydroxy acid dehydrogenase. This mutation was utilized to study the genetics and physiological expression of Had +. Had+ was mapped to the X chromosome at 54.4 and seems to be the structural gene for the enzyme. Enzyme activity in male and female flies indicates that the gene shows both dosage compensation independent from dose effect and differential activity during ontogeny. Electrophoretic mobility data indicate that the enzyme is a dimer which forms by random association of subunits. The fact that the mutant shows no detrimental effect implies that the enzyme is dispensable, at least under laboratory conditions. The biological and technical implications of this gene-enzyme system are discussed.
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  • 45
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; 6-phosphogluconate dehydrogenase ; Pgd n lethal alleles ; rescue by dietary supplements ; hexose monophosphate shunt
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The genetic rescue of Pgd n lethal alleles, accomplished by combining them with mutations lacking glucose-6-phosphate dehydrogenase activity, has led to the hypothesis that Pgd n lethality may be due to the accumulation of 6-phosphogluconate. In this article we report the rescue of Pgd n /Y males by dietary supplements (fructose and linolenate) designed to minimize 6-phosphogluconate production.
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  • 46
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    Biochemical genetics 17 (1979), S. 1-22 
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; esterase 6 ; allozymes ; biochemical properties
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Biochemical properties of esterase 6 in Drosophila melanogaster were investigated using partially purified preparations from three genotypes, 1/1, 1/2, and 2/2. The molecular weight of the enzyme is estimated to be about 90,000, and treatment with sodium dodecylsulfate cleaves the enzyme into four units with a molecular weight of about 22,000. The activity toward 28 naturally occurring esters was assayed and shown to vary considerably with substrate, the 1/1 preparation having in general higher activity than 1/2 and 2/2, which were very similar. Heat sensitivity, the effect of metal ions, and the effects of the presence or absence of an end product were also studied. The differences demonstrated between allozymes would allow considerable scope, under appropriate conditions, for differential selection to operate between genotypes.
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  • 47
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; phosphoglucomutase ; polymorphism ; enzyme kinetics
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Phosphoglucomutase (PGM) of adult stage in Drosophila melanogaster has been characterized by gel filtration, ion-exchange chromatography, and isoelectric focusing. The two common electrophoretic variants, PGMA and PGMB, differ with respect to their kinetic and stability parameters. PGMA is more thermostable than PGMB but shows the same pH optimum, equal dependence on Mg2+, and identical molecular weight. There is no significant kinetic difference between the two allozymes at the optimum pH value, but at pH 6.0 the K m value for glucose-1,6-diphosphate of PGMB is significantly higher than that of PGMA. This difference might explain the observed selective advantage of the Pgm A allele in population studies.
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  • 48
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    Biochemical genetics 35 (1997), S. 41-49 
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; null activity ; phenoloxidase
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract In Drosophila melanogaster, two new variants affecting the activity of phenoloxidase were found in natural populations at Gomel in Belorussia and at Krasnodar in Russia. Prophenoloxidases, A 1 and A 3 , in these variants had the same mobilities on native electrophoresis as the wild type. However, enzymatic activities in their activated states were much lower than in the wild type, whereas the existence of prophenoloxidase proteins was demonstrated. Egg-to-adult and relative viabilities in the variants did not decrease at temperatures between 18 and 29°C. Genetic analyses indicated that the genes showing the phenotype of variants are new alleles of Mox and Dox-3 on the second chromosome.
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  • 49
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; alcohol dehydrogenase ; enzyme biological activity ; toxicity of alcohols
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The toxicity of the first eight primary alcohols and of four secondary alcohols was compared in a wild-type strain (having active ADH) and an ADH-negative mutant. Differences between lc 50 measured in the two strains allowed an evaluation of the biological activity of the enzyme. In vitro, ADH is mainly active on secondary alcohols, while in vivo its main role is the detoxification and metabolism of ethanol. These observations suggest that originally ADH was involved in unknown metabolic pathways and that its utilization in ethanol metabolism could be a recent event.
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  • 50
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    Biochemical genetics 14 (1976), S. 237-243 
    ISSN: 1573-4927
    Keywords: null alleles ; antibody purification ; Drosophila melanogaster ; immunological methods
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Extracts from an acid phosphatase CRM− null mutant of Drosophila melanogaster were used to eliminate contaminating antibodies in a nonspecific preparation of anti-acid phosphatase serum. This method of producing specific antisera makes unnecessary the rigorous purification of an antigen prior to immunization attempts in those cases where CRM− null mutants of the antigen are available. Antisera so prepared could be used for a wide variety of purposes.
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  • 51
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    Biochemical genetics 14 (1976), S. 299-308 
    ISSN: 1573-4927
    Keywords: allozymes ; alcohol dehydrogenase ; Drosophila melanogaster
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Keeping Drosophila cultures at 28 C results in elimination of all minor multiple ADH bands, thought to be due to conformational change. Thus in diploid and triploid adults heterozygous for the Adh F and Adh Salleles, relative staining intensities are found for the three bands which were in conformity with the assumption that both alleles are equally expressed. Among all polymorphic strains derived from natural Central European and Mediterranean populations, the strain +Tüb is unique in that its Adh Fallele is closely linked to a new recessive lethal factor, named 1(2)Stm. All Adh F 1/AdhF 1 pupae are unable to emerge, and die. The lethal effect is obvious 50 hr earlier by retarded eye, bristle, and body wall pigmentation. Although all pupae of the phenotype F die, Adh F allele frequency scarcely seems to be lowered in this natural population.
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  • 52
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    Biochemical genetics 14 (1976), S. 383-387 
    ISSN: 1573-4927
    Keywords: allozymes ; thermostability ; alcohol dehydrogenase ; Drosophila melanogaster
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Two Drosophila melanogaster strains, each heterozygous for “fast” and “slow” alleles at the Adh locus, and each having balanced second chromosomes, were found to differ in the apparent thermostability of the slow allozyme. The two strains were crossed, and F1heterozygotes were separated on the basis of the origin of the slow allele. After electrophoresis, the cellulose acetate strips were treated 1 1/2 min at 35 C. The putatively more sensitive allozyme showed a strikingly greater response to heat. These findings further support the conclusion that electrophoretically cryptic allelic differences exist which are expressed in thermostability differences. Further application of this approach has revealed one similar sensitive slow allozyme and three cases of a relatively resistant fast ADH allozyme in wild-caught flies.
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  • 53
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; 6-phosphogluconolactonase ; hexose monophosphate shunt ; Pgd n Zw n mutants
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Using a double mutant strain, Pgd n Zw n , we have developed an assay for 6-phosphogluconolactonase activity and have demonstrated its occurrence in adult Drosophila melanogaster.
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  • 54
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    Biochemical genetics 17 (1979), S. 97-104 
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; form II RNA polymerase initiation sites ; chromomeres
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The in vitro incorporation of γ-32P-labeled nucleoside triphosphates into RNA by Drosophila melanogaster form II RNA polymerase from template sites which afford protection from the initiation inhibitor, polyriboinosinic acid (poly [I]), is used as a method for enumerating a specific class of transcription initiation sites on D. melanogaster DNA. Such sites number about 4000 per haploid genome for D. melanogaster. This value is in good agreement with the number of functional genetic units in the D. melanogaster genome as determined by classical cytogenetics.
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  • 55
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; Malpighian tubules ; purine transport ; eye color mutants ; riboflavin
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Uptakes of guanine into Malpighian tubules of wild-type Drosophila and the eye color mutants white (w), brown (bw), and pink-peach (p p) have been compared. Tubules for each of these mutants are unable to concentrate guanine intracellularly. The transport of xanthine and riboflavin is also deficient in w tubules. The transport of guanosine, adenine, hypoxanthine, and guanosine monophosphate is similar in wild-type and white Malpighian tubules. These data and other information about these mutants make it likely that these pteridine-deficient eye color mutants do not produce pigments because of the inability to transport a pteridine precursor. This view supports the hypothesis that mutants which lack both pteridine and ommochromes do so because precursors to both classes of pigments share a common transport system.
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  • 56
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    Molecular and cellular biochemistry 193 (1999), S. 103-108 
    ISSN: 1573-4919
    Keywords: Poly(ADP-ribose) polymerase ; Drosophila melanogaster ; alternative splicing ; apoptosis ; DNA repair ; development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Poly(ADP-ribose) polymerase (PARP) is conserved in eukaryotes. To analyze the function of PARP, we isolated and characterized the gene for PARP in Drosophila melanogaster. The PARP gene consisted of six translatable exons and spanned more than 50 kb. The DNA binding domain is encoded by exons 1-4. Although the consensus cleavage site of CED-3 like protease during apoptosis is conserved from human to Xenopus laevis PARPs, it is neither conserved in the corresponding region of Drosophila nor Sarcophaga peregrina. There are two cDNAs species in Drosophila. One cDNA could encode the full length PARP protein (PARP I), while the other is a truncated cDNA which could encode a partial-length PARP protein (PARP II), which lacks the automodification domain and is possibly produced by alternative splicing. The expression of these two forms of PARP in E. coli demonstrated that while PARP II has the catalytic NAD-binding domain and DNA-binding domain it is enzymatically inactive. On the other hand PARP I is active. A deletion mutant of PARP gene could grow to the end of embryogenesis but did not grow to the adult fly. These results suggest that the PARP gene plays an important function during the development of Drosophila.
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  • 57
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    Biochemical genetics 13 (1975), S. 263-271 
    ISSN: 1573-4927
    Keywords: allozymes ; α-glycerophosphate dehydrogenase ; Drosophila melanogaster
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract On the basis of band staining intensities in electrophoretic runs of single flies homozygous and heterozygous for two alleles at the autosomal locus for GPDH, F allele activity is believed to be 8% lower than S allele activity. Indeed, the intensity distribution in the patterns of FSS and FFS triploid females shows that both are not equally expressed. On a per fly or live weight basis, females with two and three doses of the Gpdh gene show bands with equal staining intensity, thus exhibiting a dosage effect when GPDH activity is estimated on a per cell basis.
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  • 58
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; enzyme variation ; alcohol dehydrogenase ; electrophoretically identical alleles
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A new variant of alcohol dehydrogenase (ADH 71k) was found in a laboratory stock of Drosophila melanogaster. ADH in this stock had the same electrophoretic mobility as the F variant both on acrylamide and on agar. Activity levels were similar to the levels in F flies at temperatures between 15 and 25 C. But while ADH F enzyme is inactivated rapidly at 40 C, ADH 71k is still active. Also, ADH S is not inactivated at this temperature, but has a far lower activity per fly than ADH 71k. Genetic analysis showed that the new variant is an allele of the Adh locus.
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  • 59
    ISSN: 1573-4927
    Keywords: electrophoresis ; alcohol dehydrogenase ; Drosophila melanogaster ; isozymes ; genetic variation
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    Notes: Abstract Adult Drosophila melanogaster flies collected from populations broadly dispersed over ecological and geographic strata of North Carolina, and over a period of 4 years, were analyzed for alcohol dehydrogenase phenotypes by gel electrophoresis. Gene frequencies in spring-summer-fall field collections were remarkably stable over all strata. Two winter collections exhibited contrasting gene frequency changes. In one case the results are interpreted in terms of long-distance migration from Florida, while the other is explicable by assignment of a causal role to environmental factors which accompany the winter season.
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  • 60
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    Biochemical genetics 16 (1978), S. 769-775 
    ISSN: 1573-4927
    Keywords: hidden variation ; α-GPDH ; Drosophila melanogaster
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The level of hidden variation in populations of Drosophila melanogaster at the Gpdh + locus was determined by thermal stability studies of the protein. The results indicate a lack of variation using these methods both in and between the two common electrophoretic variants. It is suggested that α-GPDH is conserved in primary structure, which may be related to its critical role in flight muscle metabolism.
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  • 61
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    Biochemical genetics 16 (1978), S. 855-865 
    ISSN: 1573-4927
    Keywords: kynurenine hydroxylase ; cinnabar locus ; EMS mutagenesis ; Drosophila melanogaster
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A study was undertaken to isolate mutations affecting the temporal appearance of kynurenine hydroxylase in Drosophila melanogaster. Such mutations, lacking or having reduced enzyme activity at the larval or pupal stage only, could represent changes in regulatory functions. Mutagenesis was carried out using EMS. Potential mutations were isolated from mass F1 cultures. The screening of large numbers of individuals was made possible by the use of the mutant red, which allowed visual classification for the presence or absence of the enzyme at both stages. From a series of six mutagenesis experiments 111,561 chromosomes were tested, and 122 phenotypically mutant F1 individuals were found. From these, 38 inheritable mutations were isolated which, by phenotypic observation, lacked or had reduced enzyme activity at the larval and pupal stages. Assay of enzyme activity levels in several of the mutants confirmed the phenotypic data. All of the 27 mutations that could be tested further are recessive and behave as cinnabar alleles. Complementation tests were performed between these 27 mutant stocks, and no complementation in the production of eye color has been seen between the mutants examined. When extended collection periods were used, a significantly higher percentage of inheritable mutations was isolated from the first 3 days of the screen. Over 80% of the F1 phenotypic mutants could be classified as mosaics, which indicates that cinnabar can be autonomous under certain conditions. The failure to isolate mutations in possible regulatory function is discussed.
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  • 62
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    Biochemical genetics 17 (1979), S. 149-158 
    ISSN: 1573-4927
    Keywords: xanthommatin synthesis ; scarlet mutants ; Drosophila melanogaster ; temperature-sensitive mutants
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Six new EMS-induced scarlet mutants were selected. Four of these were partially pigmented, with xanthommatin levels ranging from 12% to 45% of normal. In one (st 754ts), pigment production was temperature sensitive; the level of xanthommatin changed from less than 10% of normal at 29 C to more than 70% at 18 C. In all of the new mutants tested, the level of early pupal 3-hydroxykynurenine was as low as low as that in st 1. Thus reduced larval accumulation of this metabolite also appears to be a characteristic feature of scarlet mutants. Temperature-pulse and temperature-shift experiments were carried out with st 754ts to determine the temperature-sensitive period for the scarlet gene during development. The major sensitive period commenced prior to the onset of pigmentation and was over before adult emergence. Thus the initiation of xanthommatin synthesis is not brought about by the activation of the scarlet gene. In similar experiments carried out with a temperature-sensitive white mutant (w bl), a similar temperature-sensitive period was obtained.
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  • 63
    ISSN: 1573-4927
    Keywords: Drosophila melanogaster ; biopterin synthesis ; oxidation of dihydropterins
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract An enzyme which has been named “biopterin synthase” has been discovered in Drosophila melanogaster. This enzyme, which has been purified 200-fold from extracts of Drosophila, catalyzes the conversion of sepiapterin to dihydrobiopterin, or oxidized sepiapterin to biopterin. The K m values for the two substrates are 63 µm for sepiapterin and 10 µm for oxidized sepiapterin. NADPH is required in this enzymatic reaction. An analysis of enzyme activity during development in Drosophila indicates a correlation between enzyme activity and biopterin content at various development stages. Another enzyme, called “dihydropterin oxidase,” was also discovered and partially purified. This enzyme catalyzes the oxidation of dihydropterin compounds to the corresponding pterin compounds. For example, sepiapterin (a dihydropterin) is oxidized to oxidized sepiapterin in the presence of this enzyme. The only dihydropterin that has been tested that is not a substrate for this enzyme is dihydroneopterin triphosphate, the compound thought to be a precursor for all naturally occurring pterins and dihydropterins. Since the action of dihydropterin oxidase is reduced significantly when the concentration of oxygen is very low, it is likely that this enzyme uses molecular oxygen as the oxidizing agent during the oxidation of dihydropterins. Neither NAD+ or NADP+ is required. In the presence of the two enzymes dihydropterin oxidase and biopterin synthase, sepiapterin is converted to biopterin. However, in the presence of biopterin synthase alone, sepiapterin is converted to dihydrobiopterin.
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  • 64
    ISSN: 1573-3297
    Keywords: Drosophila melanogaster ; egg laying ; remating ; sperm competition
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    Topics: Biology , Psychology
    Notes: Abstract MaleDrosophila melanogaster from lines artificially selected to have divergent life histories were tested to determine if they differed, in their effects on female reproductive behavior. During the first 5 days after mating, males from short-generation populations caused females to lay eggs at a faster rate than did males from long-generation populations. This faster oviposition rate resulted in greater numbers of adult progeny produced by short-generation males. During the period 6–21 days after mating, long-generation males fathered more adult progeny. Females that were first mated to short-generation males were more likely to remate than were females first mated to long-generation males. Rematings were interrupted in order to prevent transfer of second-male accessory fluid and sperm. Females that were first mated to long-generation males produced more progeny after interrupted matings than did females that were first mated to short-generation males.
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  • 65
    ISSN: 1573-3297
    Keywords: Drosophila melanogaster ; vision ; larvae
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    Topics: Biology , Psychology
    Notes: Abstract Larvae from seven laboratory strains and eight isofemale lines ofDrosophila melanogaster differ significantly with regard to their responses to light in a photokinesis assay in which the larvae are tested en masse. Larvae from the CA-2 laboratorystock fail to disperse on assay plates, although observations of individual CA-2 larvae suggest that the larvae are repelled by light. Larvae from all of the other laboratory stocks and all of the isofemale lines (except LI2 and NC5) avoid light in the photokinesis assay. Larvae from some stocks are much more strongly repelled by light than larvae from other stocks. LI2 larvae are unresponsive to light in most replicates of the photokinesis assay, while NC5 larvae are consistently unresponsive to light. Observations of F1 heterozygotes suggest that the allele(s) that affects the vision of LI2 and NC5 larvae has net effects on the animals' behavior that are partially dominant and recessive, respectively.
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  • 66
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    Molecular biology reports 24 (1997), S. 133-138 
    ISSN: 1573-4978
    Keywords: proteasome ; multicatalytic proteinase ; subunit composition ; insect ; crustacean ; Drosophila melanogaster ; lobster ; muscle ; Homarus americanus ; Manduca sexta
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    Biochemical genetics 34 (1996), S. 45-59 
    ISSN: 1573-4927
    Keywords: RNA polymerase II ; Ultrabithorax ; transcription ; α-amanitin ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Interference between different classes of RNA polymerase II alleles causes a mutant phenotype called the “Ubx effect” that resembles one seen in flies haploinsufficient for the transcription factor,Ultrabithorax (Ubx). Flies carrying the mutation in the largest subunit ofDrosophila RNA polymerase II,RpII215 4, display the Ubx effect when heterozygous as inRpII215 4/+ but not when homozygous mutant or wild type. In this report we demonstrate that the interaction between alleles in different classes of polymerase occurs even in the absence of transcription by the wild-type polymerase. We utilized the resistance to the transcriptional inhibitor α-amanitin conferred byRpII215 4 to show thatRpII215 4/+ flies raised on α-amanitin-containing food still show the Ubx effect and are indistinguishable from flies raised on normal food. We demonstrate using HPLC that the intracellular concentration of α-amanitin in the developing larvae is sufficient to inhibit transcription by α-amanitin-sensitive polymerase. Furthermore, fluorescein-labeled α-amanitin accumulates in imaginal discs, which are the precursor cells for the tissue showing the homeotic transformation in adults. We conclude that the interaction between different classes of RNA polymerase II alleles resulting in the Ubx effect occurs prior to the block in transcription caused by α-amanitin.
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    Biochemical genetics 34 (1996), S. 45-59 
    ISSN: 1573-4927
    Keywords: RNA polymerase II ; Ultrabithorax ; transcription ; α-amanitin ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Interference between different classes of RNA polymerase II alleles causes a mutant phenotype called the “Ubx effect” that resembles one seen in flies haploinsufficient for the transcription factor,Ultrabithorax (Ubx). Flies carrying the mutation in the largest subunit ofDrosophila RNA polymerase II,RpII215 4, display the Ubx effect when heterozygous as inRpII215 4/+ but not when homozygous mutant or wild type. In this report we demonstrate that the interaction between alleles in different classes of polymerase occurs even in the absence of transcription by the wild-type polymerase. We utilized the resistance to the transcriptional inhibitor α-amanitin conferred byRpII215 4 to show thatRpII215 4/+ flies raised on α-amanitin-containing food still show the Ubx effect and are indistinguishable from flies raised on normal food. We demonstrate using HPLC that the intracellular concentration of α-amanitin in the developing larvae is sufficient to inhibit transcription by α-amanitin-sensitive polymerase. Furthermore, fluorescein-labeled α-amanitin accumulates in imaginal discs, which are the precursor cells for the tissue showing the homeotic transformation in adults. We conclude that the interaction between different classes of RNA polymerase II alleles resulting in the Ubx effect occurs prior to the block in transcription caused by α-amanitin.
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  • 69
    ISSN: 1573-4927
    Keywords: α-glucosidases ; Drosophila melanogaster ; carbohydrate metabolism
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract InbredDrosophila melanogaster stocks were surveyed for α-glucosidases with nondenaturing gel electrophoresis using a fluorogenic substrate to stain the gels. The glucosidase most active under these conditions is polymorphic. We established that the polymorphism is genetic in origin and that the glucosidase was not likely to be a previously characterized enzyme. The gene encoding the enzyme was mapped cytogenetically to 33 A1-2- 33A8-B1, confirming that this is an enzyme not yet reported inD. melanogaster. The enzyme was partially purified by elution from nondenaturing gels, which enabled us to establish that it has optimal activity at pH 6 and interacts most strongly with α-1–4 glucosides. A developmental and tissue survey suggested that this enzyme could have a purely digestive role or be involved in carbohydrate metabolism inside the organism. We propose that this enzyme is involved in either starch digestion or glycogen metabolism.
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  • 70
    ISSN: 1573-4927
    Keywords: α-glucosidases ; Drosophila melanogaster ; carbohydrate metabolism
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract InbredDrosophila melanogaster stocks were surveyed for α-glucosidases with nondenaturing gel electrophoresis using a fluorogenic substrate to stain the gels. The glucosidase most active under these conditions is polymorphic. We established that the polymorphism is genetic in origin and that the glucosidase was not likely to be a previously characterized enzyme. The gene encoding the enzyme was mapped cytogenetically to 33 A1-2- 33A8-B1, confirming that this is an enzyme not yet reported inD. melanogaster. The enzyme was partially purified by elution from nondenaturing gels, which enabled us to establish that it has optimal activity at pH 6 and interacts most strongly with α-1–4 glucosides. A developmental and tissue survey suggested that this enzyme could have a purely digestive role or be involved in carbohydrate metabolism inside the organism. We propose that this enzyme is involved in either starch digestion or glycogen metabolism.
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    Theoretical and applied genetics 55 (1979), S. 231-238 
    ISSN: 1432-2242
    Keywords: Drosophila melanogaster ; Scute locus ; Maps ; Operon-like model
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The functional expression of 12 scute alleles in homozygotes and compounds of Drosophila melanogaster at 14°, 22°, 30°C is analysed. Based on the data obtained, linear maps for bristles and mutations are built. The basic features of the maps, clustering and polarity, are invariable with respect to temperature, scute gene dosage and cross direction. In addition local dominance of the norm over bristle reduction was produced by the scute mutation; different types of complementation reactions were established for each bristle. The gene scute is treated as an operon-like system, composed of 3–4 cistrons with each controlling the formation of bristles on a particular region of the fly's body. This model argues well with the structure of maps constructed and implies a post-translational level of initial events of bristle-formation process.
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    Theoretical and applied genetics 91 (1995), S. 1095-1100 
    ISSN: 1432-2242
    Keywords: Drosophila melanogaster ; Scute locus ; Penetrance ; Polygenes ; Transposable elements
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We wanted to determine whether there is a correlation between the quantitative character, the penetrance of the loss of humeral bristles in scute lines, and the distribution of transposable genetic elements in their genomes. We derived 18 isogenic lines with penetrance ranging between 2.8% and 92.0% from six mutant lines. The localization of the transposable elements (TEs) P, mdg1, Dm412, copia, gypsy and B104 was determined in all isogenic derivatives by in situ hybridization. The total number of the TE sites over all lines was 180. A comparison of the distribution of the TEs in the isogenic lines revealed the location of sites typical of lines with similar penetrance, no matter which parental line was involved. The results obtained suggest that such typical sites appear to tag the genome regions where the polygenes affecting the character in question are most likely to be found.
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  • 73
    ISSN: 1617-4623
    Keywords: Drosophila melanogaster ; Hybrid ; dysgenesis ; I factor ; Transposition ; Complementation
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    Topics: Biology
    Notes: Abstract Non-LTR retrotransposons, also known as LINEs, transpose by reverse transcription of an RNA intermediate. Their mechanism of transposition is apparently different from that of retrotransposons and similar to that of proviruses of retroviruses. The I factor is responsible for the I-R system of hybrid dysgenesis inDrosophila melanogaster. Inducer strains contain several functional I factors whereas reactive strains do not. Transposition of I factors can be experimentally induced: they are stable in inducer strains, but transpose at high frequency in the germline of females, known as SF females, produced by crossing reactive females and inducer males. We have constructed an I element, calledIviP2, marked with thevermilion gene, the coding sequence of which was interrupted by an intron. Splicing of the intron can only occur in the transcript initiated from the I element promoter. Transposed copies expressing a wild-typevermilion phenotype were recovered in the germline of SF females in which I factors were actively transposing. This indicates thattrans-complementation of a defective I element, deficient for the second open reading frame, by functional I factors can occur in the germline of dysgenic females.
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    Molecular genetics and genomics 249 (1995), S. 673-681 
    ISSN: 1617-4623
    Keywords: Spontaneous mutation ; Transposable elements ; Drosophila melanogaster ; scarlet gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Six spontaneous mutations of the scarlet (st) locus of Drosophila melanogaster have been studied at the molecular level. Two of the mutants (st 1 and st sp) arose in laboratory populations, while the other four (st cob, st ct89, st dct and st dv) were isolated from natural populations. In five of these there is a DNA insertion within the st region and in four cases the insertion has been identified as being a transposable element; these include the retrotransposons 412 and B104/roo, and also jockey a member of the LINE family. In the other case (st dct), the insertion appears to consist of partially duplicated st sequences. In two of the mutants (st 1 and st dv) the same transposable element (412) has inserted in the same orientation at exactly the same site within the st gene. The transposable element insertions are found in intron and exon regions of the st gene and also in the putative upstream regulatory region; insertions located in introns or exons result in the production of truncated st transcripts. The results show that the same types of transposable elements that cause spontaneous mutation in laboratory stocks of D. melanogaster also cause mutation in the wild.
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  • 75
    ISSN: 1617-4623
    Keywords: Cytochrome P450 genes ; Drosophila melanogaster ; Gene clusters ; Multigene family ; Insecticide resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Twelve cytochrome P450 cDNA fragments were cloned fromDrosophila melanogaster by reverse transcriptase/PCR (RT/PCR) using degenerate oligonucleotide primers. The corresponding genes belong to several subfamilies of the CYP4 and CYP9 P450 families. Only two of these genes,Cyp4d1 andCyp4d2, have previously been described.In situ hybridization of each of the cDNA fragments showed two clusters of genes; one near the tip of theX chromosome and the other on the left arm of chromosome2. Interestingly the latter cluster comprises widely divergent genes belonging both to the CYP9 and CYP4 families and also to the CYP6 family (Cyp6a2). Putative allelic variants of several of the genes were found in different insecticide-resistant and -susceptible strains (Hikone R, Haag 79 and Oregon R). The identification of these genes and alleles will allow us to clarify the involvement of P450s in xenobiotic metabolism and will facilitate a genetic analysis of P450 functions in insects.
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  • 76
    ISSN: 1617-4623
    Keywords: Key wordspara ; Voltage-gated sodium channel ; Insecticide resistance ; DDT/pyrethroids ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The gene para in Drosophila melanogaster encodes an α subunit of voltage-activated sodium channels, the presumed site of action of DDT and pyrethroid insecticides. We used an existing collection of Drosophila para mutants to examine the molecular basis of target-site resistance to pyrethroids and DDT. Six out of thirteen mutants tested were associated with a largely dominant, 10- to 30-fold increase in DDT resistance. The amino acid lesions associated with these alleles defined four sites in the sodium channel polypeptide where a mutational change can cause resistance: within the intracellular loop between S4 and S5 in homology domains I and III, within the pore region of homology domain III, and within S6 in homology domain III. Some of these sites are analogous with those defined by knockdown resistance (kdr) and super-kdr resistance-associated mutations in houseflies and other insects, but are located in different homologous units of the channel polypeptide. We find a striking synergism in resistance levels with particular heterozygous combinations of para alleles that appears to mimic the super-kdr double mutant housefly phenotype. Our results indicate that the alleles analyzed from natural populations represent only a subset of mutations that can confer resistance. The implications for the binding site of pyrethroids and mechanisms of target-site insensitivity are discussed.
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  • 77
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    Molecular genetics and genomics 256 (1997), S. 652-660 
    ISSN: 1617-4623
    Keywords: Key words S-adenosylmethionine decarboxylase ; Polyamine synthesis ; Drosophila melanogaster
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    Topics: Biology
    Notes: Abstract S-adenosylmethionine decarboxylase is a key enzyme in the synthesis of polyamines. These small cationic molecules are required for growth and development in all organisms. A wealth of biological processes, including synthesis of DNA and protein and condensation of chromatin, involve polyamines. Inhibition of polyamine synthesis has been proposed for treatment of cancer but this requires more knowledge about the in vivo function of polyamines. We report here the cloning of the S-adenosylmethionine decarboxylase gene from Drosophila melanogaster and the analysis of corresponding mutants. The mutant phenotypes are similar to those previously described for ribosomal protein genes (Minutes) and rRNA genes (bobbed ). This work elucidates the in vivo consequences of impaired polyamine synthesis with respect to the development of a whole animal.
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  • 78
    ISSN: 1617-4623
    Keywords: Key words Proteasome ; Apoptosis ; sug-1 ; Drosophila melanogaster ; Nervous system
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    Notes: Abstract The proteasome plays essential roles in a variety of cellular processes, including degradation of the bulk of cellular proteins, degradation of short-lived proteins such as cell cycle regulators, generation of antigenic peptides, and mediating programmed cell death. One of the best characterized subunits of the 26S proteasome is encoded by the yeast gene SUG1. We report here the cloning and characterization of the Drosophila homolog of this gene, Pros45. At the protein level, Pros45 is highly conserved with respect to its homologs in a variety of taxa: it shows 74% identity to yeast Sug1; 86% to mouse m56/mSug1/FZA-B; 87% to human Trip1; and 97% to moth 18-56. Using a genomic clone as a probe for in situ hyridization to polytene chromesomes, we demonstrated that Pros45 maps to 19F, near the base of the X chromosome. Use of a pros45 cDNA clone as a probe revealed a second site of hybridization at 99CD. Pros45 mRNA is found in the unfertilized egg and in all cells of the early embryo. By the end of embryogenesis, Pros45 is expressed predominantly in the central nervous system. Targeted expression of Pros45 in a variety of different cells using the Gal4 UAS P-element system failed to generate an overt phenotype. This study provides the foundation for further examination of the role of the 26S proteasome in homeostasis and development in Drosophila.
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  • 79
    ISSN: 1617-4623
    Keywords: Key words NADH:ubiquinone oxidoreductase acyl carrier protein ; Drosophila melanogaster ; Alternative splicing ; P-element-induced mutation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have isolated the Drosophila melanogaster gene encoding the mitochondrial acyl carrier protein (mtACP), a subunit of NADH:ubiquinone oxidoreductase involved in de novo fatty acid synthesis in the mitochondrion. This gene expresses two distinct mature transcripts by alternative splicing, which encode mature polypeptides of 86 (mtACP1A) and 88 (mtACP1B) amino acids, respectively. Drosophila mtACP1 is 72% identical to mammalian mtACP, 47% identical to Arabidopsis thaliana mtACP, and 46% identical to Neurospora crassa mtACP. The most highly conserved region encompasses the site that binds pantetheine-4′-phosphate in all known ACPs. Southern analysis of genomic DNA and in situ hybridization to salivary gland chromosomes indicate that a single gene (mtacp1), located at 61F6–8, encodes the two isoforms of D. melanogaster mtACP1. Sequence analysis revealed that the gene contains four exons and that exons IIIA and IIIB are alternatively spliced. A P-element-induced loss-of-function mutation in the mtacp1 gene causes lethality, indicating that the gene is essential for viability. Developmental Northern analysis shows that mtacp1 is expressed at higher levels during late embryogenesis, in the pupa and in the adult. RNA in situ hybridization on embryos indicates that the mtacp1 gene is highly expressed in the tracheal system. Zygotic mtacp1 function is required for both male and female gametogenesis.
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  • 80
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    Molecular genetics and genomics 250 (1996), S. 601-613 
    ISSN: 1617-4623
    Keywords: Key words Sex-influenced modifier ; Position effect variegation ; Dosage effects ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Modifier of white (Mow), a dominant trans-acting gene, has been identified through a mutagenic screen for second-site loci that alter the level of expression of the white eye color locus. Mow reduces the expression of white in most developmental stages, but enhances its expression in the pupal stage, the time at which the major contribution to the adult phenotype is made. Tests with an Alcohol dehydrogenase promoter-white reporter and a series of white truncation constructs have shown that Mow fails to affect the reporter; cis-regulatory mutations of white also show no response, suggesting a requirement for white regulatory domains for interaction with Mow. A quantitative analysis of steady-state transcript levels reveals that the white mRNA level decreases in the presence of one dose of Mow in larvae and adults, but the reduction is greater in females than males. Two other functionally related genes, brown and scarlet, also exhibit a similar sexually dimorphic alteration in expression, mediated by Mow. In the mid-pupal stage, by contrast, the level of white and brown mRNA is increased by Mow. In addition, Mow acts as a weak suppressor of position effect variegation (PEV). These observations suggest a connection between dosage modulation of gene expression and suppression of position-effect variegation.
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  • 81
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    Molecular genetics and genomics 251 (1996), S. 130-138 
    ISSN: 1617-4623
    Keywords: Transposable elements ; Drosophila melanogaster ; Quantitative variation ; Population genetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mobilization rates of nine families of transposable elements (P, hobo, FB, gypsy, 412, copia, blood, 297, andjockey) were estimated by using 182 lines. Lines were started from a completely isogenic population ofDrosophila melanogaster, carrying the markersepia as an indicator of possible contamination, and have been accumulating spontaneous mutations independently for 80 generations of brother-sister (or two double-first-cousin) matings. Transposable element movements have been analyzed in complete genomes by the Southern technique. Mobilization was a rare event, with an average rate of 10−5 per site per generation. The most active element wasFB. In contrast, the retroelementsgypsy andblood did not move at all. Most changes in restriction patterns were consistent with rearrangements rather than with true transposition. The euchromatic or heterochromatic location of elements was tested by comparing insertion patterns from adults and salivary glands. Certain putative rearrangements involved heterochromatic copies of the retroelements412, copia or297. Clustering of movement across families was observed, suggesting that movement of different families may be non-independent. An association between modified insertion patterns and mutant effects on quantitative traits shows that spontaneous transposition events cause continuous variation.
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  • 82
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    Molecular genetics and genomics 251 (1996), S. 130-138 
    ISSN: 1617-4623
    Keywords: Key words Transposable elements ; Drosophila melanogaster ; Quantitative variation ; Population genetics
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    Notes: Abstract  Mobilization rates of nine families of transposable elements (P, hobo, FB, gypsy, 412, copia, blood, 297, and jockey) were estimated by using 182 lines. Lines were started from a completely isogenic population of Drosophila melanogaster, carrying the marker sepia as an indicator of possible contamination, and have been accumulating spontaneous mutations independently for 80 generations of brother-sister (or two double-first-cousin) matings. Transposable element movements have been analyzed in complete genomes by the Southern technique. Mobilization was a rare event, with an average rate of 10-5 per site per generation. The most active element was FB. In contrast, the retroelements gypsy and blood did not move at all. Most changes in restriction patterns were consistent with rearrangements rather than with true transposition. The euchromatic or heterochromatic location of elements was tested by comparing insertion patterns from adults and salivary glands. Certain putative rearrangements involved heterochromatic copies of the retroelements 412, copia or 297. Clustering of movement across families was observed, suggesting that movement of different families may be non-independent. An association between modified insertion patterns and mutant effects on quantitative traits shows that spontaneous transposition events cause continuous variation.
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  • 83
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    Molecular genetics and genomics 258 (1998), S. 457-465 
    ISSN: 1617-4623
    Keywords: Key words Adaptive mutation ; Drosophila melanogaster ; Mutagenesis ; RNA polymerase II ; Suppressor selection
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    Topics: Biology
    Notes: Abstract We used a mutagenesis and selection procedure in Drosophila melanogaster to recover rare allele-specific suppressor mutations. More than 11 million flies mutant for one of five recessive-lethal mutations in the two largest subunits of RNA polymerase II were selected for additional mutations that restored viability. Forty-one suppressor mutations were recovered. At least 16 are extragenic, identifying a minimum of three loci, two of which do not map near genes known to encode subunits of RNA polymerase II. At most, 25 are intragenic, 4 reverting the initial altered nucleotide back to wild type. Sequence analysis of interacting mutations in the two largest subunits identified a discrete domain in each subunit. These domains might be contact points for the subunits. Finally, our selections were large enough to allow recovery of multiple independent changes in the same nucleotides yet mutations in other equally likely targets were not recovered. The mutations recovered are not random and might provide insights into possible mechanisms for mutagenesis in eukaryotes.
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  • 84
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    Neuroscience and behavioral physiology 27 (1997), S. 254-257 
    ISSN: 1573-899X
    Keywords: Drosophila melanogaster ; P-element ; operative learning ; dunce mutant ; rutabaga mutant
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    Topics: Biology , Medicine
    Notes: Abstract A collection ofDrosophila melanogaster mutants was created by insertion of a P-element into autosomes at a rate of one copy per genome. The abilities of 64 homozygous P-insertion mutants to produce a form of associative behavior were determined. Testing was based on an original paradigm of operant learning: interactions betweenDrosophila individuals when placed in a group situation in which the flies learned to inhibit their own activity to avoid punishment in the form of conflict with other individuals. Four lines were found in which, like the known learning mutants dunce and rutabaga, individuals did not show changes in their initial responses to each other. These lines were also studied using other paradigms of associative learning.
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  • 85
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    Evolutionary ecology 12 (1998), S. 363-376 
    ISSN: 1573-8477
    Keywords: adult crowding ; density-dependent selection ; Drosophila melanogaster ; life-history
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    Notes: Abstract The effects of adult crowding on two components of fitness were studied in three sets of Drosophila melanogaster populations, subjected to life-stage-specific, density-dependent natural selection in the laboratory for over 50 generations. Three days of crowding, early in adult life, were observed to increase mortality significantly during the episode of crowding, as well as decrease subsequent fecundity. Populations selected for adaptation to high adult densities suffered significantly lower mortality during episodes of adult crowding, as compared to populations selected specifically for adaptation to larval crowding, as well as control populations typically maintained at low larval and adult densities. Moreover, populations adapted to larval crowding seemed to be adversely affected by adult crowding to a greater extent than the controls, raising the possibility of trade-offs between adaptations to larval and adult crowding, respectively. Preliminary evidence suggests that the populations adapted to adult crowding may have evolved a propensity to stay away from the food medium, which is where most deaths occur when adults are crowded in culture vials.
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  • 86
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    Behavior genetics 28 (1998), S. 137-151 
    ISSN: 1573-3297
    Keywords: Flightless mutants ; wing-beat frequency ; wing size ; body size ; mating behavior ; courtship song ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Psychology
    Notes: Abstract Flightless mutations that affect wing-beat frequency (WBF) of Drosophila melanogaster were examined for their effect on male courtship. WBFs were measured using a fixed-wire tether that completely supports the fly in an attitude similar to hovering flight. The two spontaneous mutations, one of which reduces WBF to one-half normal and the other to zero, were placed on an isogenic background and were compared to an isogenic wildtype strain and to a genetically heterogeneous wildtype strain. Time to mating under noncompetitive conditions (single pair matings) was not significantly different among the four male types in one experiment. In a second experiment, although the time to mating varied significantly among the four male types, there was no association between the WBF that was characteristic of a male type and the length of time to mating. Time to mating was not significantly correlated with WBF, wing size, or body mass in either experiment. Genetically heterogeneous wild-type females were significantly more receptive (had shorter times to mating) than inbred wild-type females toward all four male types. During the time-to-mating tests, all four male types appeared to show typical courtship behaviors. Therefore, the male types were compared for possible differences in four components of the male courtship song: sine song frequency, interpulse interval, intrapulse frequency (=carrier frequency), and wing cycles per pulse. One or another of these components showed significant differences among the four male types (e.g., genetically heterogeneous, wild-type males showed a significantly higher sine song frequency and intrapulse frequency than males of the three isogenic types). However, the average values for all four male types were within reported wild-type ranges for each courtship song component. Although the two mutations drastically reduce WBF during tethered flight, they do not have any similar major effect on courtship. Apparently they affect muscles or neuronal control mechanisms that are not common elements shared by the wing movement of flight and male courtship song.
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  • 87
    ISSN: 1573-3297
    Keywords: Drosophila melanogaster ; phenylthiocarbamide ; isogenic strain ; oviposition ; F1 cross ; backcross ; F2 cross
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    Topics: Biology , Psychology
    Notes: Abstract Seven isogenic strains of Drosophila melanogaster were assayed for oviposition preference on food with phenylthiocarbamide (PTC) versus plain food. There was significant variation among strains for the percentage of eggs oviposited on each medium, ranging from 70±4% (SE) preference for plain food to no significant preference. Reciprocal hybrid, backcross, and F2 generations derived from two extreme parent strains revealed significant additive and nonadditive genetic variation but no evidence of maternal, paternal, or sex-chromosome effects.
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  • 88
    ISSN: 1573-6857
    Keywords: Drosophila melanogaster ; P elements ; population studies ; transposable elements
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    Topics: Biology
    Notes: Abstract The use of transposable elements (TEs) as genetic drive mechanisms was explored using Drosophila melanogaster as a model system. Alternative strategies, employing autonomous and nonautonomous P element constructs were compared for their efficiency in driving the ry+ allele into populations homozygous for a ry- allele at the genomic rosy locus. Transformed flies were introduced at 1%, 5%, and 10% starting frequencies to establish a series of populations that were monitored over the course of 40 generations, using both phenotypic and molecular assays. The transposon-borne ry+ marker allele spread rapidly in almost all populations when introduced at 5% and 10% seed frequencies, but 1% introductions frequently failed to become established. A similar initial rapid increase in frequency of the ry+ transposon occurred in several control populations lacking a source of transposase. Constructs carrying ry+ markers also increased to moderate frequencies in the absence of selection on the marker. The results of Southern and in situ hybridization studies indicated a strong inverse relationship between the degree of conservation of construct integrity and transposition frequency. These finding have relevance to possible future applications of transposons as genetic drive mechanisms.
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  • 89
    ISSN: 1573-6857
    Keywords: Drosophila melanogaster ; chromosomal ; inversion polymorphism ; temporal variation
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    Notes: Abstract Investigations on the chromosomal inversion polymorphism were conducted on a Korean (Taenung) natural population of D. melanogaster during the period 1978 to 1992. A total of 66 different endemic and cosmopolitan inversions were found on both major chromosome pairs II and III. Some of them proved to be rare cosmopolitan types (2LKA, 2LNS, 2LF, 2RCy, 3LM, 3RKI, and 3RK), while others were endemics. The distribution of breakpoints for endemic and rare cosmopolitan inversions are not random along the two autosome arms.With respect to frequency changes, the 15-year survey revealed that five of the cosmopolitan types (2Lt, 2RNS, 3LP, 3RC, and 3RMo) exhibit cyclical frequency changes, whereas gene arrangement 3RP shows relatively stable frequencies. Tests for correlations between gene arrangement frequencies and several climatic variables gave no clear evidence for such relationships. Only one correlation coefficient out of 64 was statistically significant.
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  • 90
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    Chromosome research 3 (1995), S. 239-244 
    ISSN: 1573-6849
    Keywords: atomic force microscope ; Drosophila melanogaster ; polytene chromosomes
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    Topics: Biology
    Notes: Abstract Polytene chromosomes from the salivary gland cells of Drosophila melanogaster were examined by atomic force microscopy. The atomic force microscope (AFM) was capable of resolving chromosomal features down to the limits of the tip sharpness, about 500 Å for pyramidal-shaped tips. Resolution was increased to 300 Å by using electron beam deposited (EBD) tips with high aspect ratios. This significantly exceeds the resolution obtainable with conventional optical microscopes, but at the cost of compromising the structural integrity of the sample. A reasonable compromise was achieved by using oxide-sharpened tips. In this case high resolution was obtained without sample degradation, but when desired these tips were also capable of sample disintegration with increased scanning force and rate. Thus, oxide-sharpened tips were used to precisely dissect defined chromosomal regions to illustrate their potential use in genetic mapping efforts. This study illustrates the utility of the AFM in the characterization and manipulation of chromosomes and chromosomal DNA.
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  • 91
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    Chromosome research 3 (1995), S. 351-360 
    ISSN: 1573-6849
    Keywords: chromatin ; Drosophila melanogaster ; Polycomb ; silencing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract InDrosophila the Polycomb group (Pc-G) proteins are responsible for the stable and heritable silencing of genes. The Pc-G apparently uses heterochromatin-like mechanisms to transcriptionally inactivate developmental regulators such as the homeotic genes. The Polycomb (Pc) protein is part of a large multimeric complex composed of other members of the Pc-G. We have identified functionally relevant domains of the Pc protein by sequencing differentPc alleles. Additionally, using a Pc-βgal fusion protein with deleted internal histidine repeats, we found that this mutant protein cannot bind to four particular target loci, but otherwise does not change the remaining overall binding pattern. We show that, in contrast to the dotted subnuclear localization of the wild-type protein, the nuclear distribution of mutant proteins becomes homogeneous. Surprisingly, inPc mutants the polyhomeotic protein, another member of the Pc-G, is also redistributed in the nucleus. Our results indicate that the appropriate subnuclear localization of the two proteins is critical for the silencing function of the Pc-G complex.
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  • 92
    ISSN: 1573-6857
    Keywords: fitness ; Drosophila melanogaster ; locomotion ; mating activity ; P DNA elements
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    Topics: Biology
    Notes: Abstract Some transposable DNA elements in higher organisms are active in somatic cells, as well as in germinal cells. What effect does the movement of DNA elements in somatic cells have on life history traits? It has previously been reported that somatically active P and mariner elements in Drosophila induce genetic damage and significantly reduce lifespan. In this study, we report that the movement of P elements in somatic cells also significantly reduces fitness, mating activity, and locomotion of Drosophila melanogaster. If other elements cause similar changes in life history traits, it is doubtful if transposable DNA elements remain active for long in somatic cells in natural populations.
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  • 93
    ISSN: 1573-8477
    Keywords: geographical variation ; encapsulation ability ; Drosophila melanogaster ; parasitoid ; Asobara tabida ; Leptopilina boulardi ; parasitoid specificity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The ability ofDrosophila melanogaster larvae in Europe to encapsulate the eggs of their most important parasitoids,Asobara tabida andLeptopilina boulardi, shows a large amount of geographical variation. Interestingly, encapsulation ability againstA. tabida is not correlated with encapsulation ability againstL. boulardi. This indicates that the encapsulation system ofD. melanogaster larvae has parasitoid-specific components. The variation in encapsulation ability can only partly be explained by the incidence of parasitism on the larvae. This means that factors other than the two parasitoid species must be selection pressures on the encapsulation ability ofD. melanogaster larvae.
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  • 94
    ISSN: 1617-4623
    Keywords: Drosophila melanogaster ; Polycomb group genes ; Tumour suppressor genes ; Homeosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genetic analysis of the 8D3;8D8-9 segment of the Drosophila melanogaster X chromosome has assigned seven complementation groups to this region, three of which are new. A Polycomb group (Pc-G) gene, multi sex combs (mxc), is characterized and mutant alleles are described. Besides common homeotic transformations characteristic of Pc-G mutants that mimic the ectopic gain of function of BX-C and ANT-C genes, mxc mutants show other phenotypes: they zygotically mimic, in males and females, the characteristic lack of germ line seen in progeny of some maternal effect mutants of the so-called posterior group (the grandchildless phenotype). Loss of normal mxc function can promote uncontrolled malignant growth which indicates a possible relationship between Pc-G genes and tumour suppressor genes. We propose that gain-of-function of genes normally repressed by the wild-type mxc product could, in mxc mutants, give rise to an incoherent signal which would be devoid of meaning in normal development. Such a signal could divert somatic and germ line developmental pathways, provoke the loss of cell affinities, but allow or promote growth.
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  • 95
    ISSN: 1617-4623
    Keywords: Key words Yolk protein ; Ovarian enhancer 1 ; Enhancer-binding factors ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  It has been reported that three different DNA regions – the fat body enhancer and ovarian enhancers 1 and 2 – direct the tissue-specific expression of yp1 and yp2 in Drosophila melanogaster. In the present study, we identified ovarian enhancer 1-specific binding proteins. Electrophoretic mobility shift assay revealed that these proteins are present in the adult ovary, but not in adult testis or fat body. Southwestern blot analysis showed that about 130 kDa and 40 kDa proteins, designated OEF1 and OEF2, respectively, from ovarian nuclear or crude extracts bind specifically to the ovarian enhancer 1. The two proteins were partially purified by streptavidin/agarose-DNA affinity chromatography, and their binding activity was confirmed by electrophoretic mobility shift assay. These ovarian enhancer factors may play an important role in the regulation of transcription of yp1 and yp2 in the ovary.
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  • 96
    ISSN: 1617-4623
    Keywords: Spontaneous mutations ; Natural populations ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A large proportion of spontaneous mutations inDrosophila melanogaster strains of laboratory origin are associated with insertions of mobile DNA elements. As a first step toward determining whether spontaneous laboratory mutations are predictive for mutational events occurring in the wild, recessivebrown (bw) eye color mutants were isolated. By inbreeding the progeny of wild-caughtDrosophila melanogaster females,bw mutations were isolated from seven separate geographic sites distributed among Japan, California, Siberia and Hungary. Among a total of 14 mutations studied, no case of transposon mutagenesis was found. At least 4 mutations are associated with small deletions in thebw gene. The remainder are inseparable from wild-typebw by Southern analysis and are presumed to be basepair changes or very small indels. Although only two spontaneousbw mutants of laboratory origin have been analyzed molecularly, one is a mobile element insertion.
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  • 97
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    Molecular genetics and genomics 247 (1995), S. 399-408 
    ISSN: 1617-4623
    Keywords: hobo element ; Transposon excision ; Transposable elements ; Drosophila melanogaster ; Drosophilidae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mobility of the hobo transposable element was determined for several strains of Drosophila melanogaster and several Drosophila species. Mobility was assessed by use of an in vivo transient assay in the soma of developing embryos, which monitored hobo excision from injected indicator plasmids. Excision was detected in a D. melanogaster strain (cn; ry 42) devoid of endogenous hobo elements only after co-injection of a helper plasmid containing functional hobo transposase under either heat shock or normal promoter regulation. Excision was also detected in D. melanogaster without helper in strains known to contain genomic copies of hobo. In Drosophila species confirmed not to contain hobo, hobo excision occurred at significant rates both in the presence and absence of co-injected helper plasmid. In four of the seven species tested, excision frequencies were two- to fivefold lower in the presence of plasmid-borne hobo. hobo excision donor sites were sequenced in indicator plasmids extracted from D. melanogaster cn; ry 42 and D. virilis embryos. In the presence of hobo transposase, the predominant excision sites were identical in both species, having breakpoints at the hobo termini with an inverted duplication of proximal insertion site DNA. However, in the absence of hobo transposase in D. virilis, excision breakpoints were apparently random and occurred distal to the hobo termini. The data indicate that hobo is capable of functioning in the soma during embryogenesis, and that its mobility is unrestricted in drosophilids. Furthermore, drosophilids not containing hobo are able to mobilize hobo, presumably by a hobo-related cross-mobilizing system. The cross-mobilizing system in D. virilis is not functionally identical to hobo with respect to excision sequence specificity.
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  • 98
    ISSN: 1617-4623
    Keywords: Drosophila melanogaster ; Larval serum protein-2 gene ; Ecdysone response element ; Ecdysone receptor ; USP/Drosophila retinoid-X receptor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Larval serum protein-2 gene (Lsp-2) of Drosophila melanogaster is uniquely expressed in the fat body tissue from the beginning of the third instar to the end of adult life. Accumulation of the larval Lsp-2 transcript is enhanced by 20-hydroxyecdysone. To study the molecular basis for ecdysone regulated Lsp-2 activity, deletion mutants of the Lsp-2 5′-flanking region were constructed by fusion to either the Escherichia coli chloramphenicol acetyltransferase (CAT) gene or to an hsp70-lacZ hybrid gene encoding β-galactosidase. Constructs transfected into Drosophila S2/M3 cells were shown to confer transient ecdysone inducibility on the reporter genes. A single functional ecdysone response element (EIRE) was localized at position — 75 relative to the Lsp-2 transcription initiation site. In gel mobility shift assays using fat body nuclear extracts or nuclear receptors synthesized in vitro, a 27-bp sequence harboring the EcRE bound both the Drosophila ecdysone receptor and the Drosophila retinoid-X homologue, Ultraspiracle, in a cooperative manner. Competition experiments indicate that the affinity of the Lsp-2 EcRE for the ecdysone receptor complex is comparable to that of the canonical EcRE of the hsp27 gene and is at least 4-fold greater than that of Fbp1, another fat body-specific Drosophila gene. Our results suggest that structural features of this EcRE determine its ability to induce ecdysone responsiveness at a lower ligand concentration and may form the basis for differential hormone responsiveness within the fat body.
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    Molecular genetics and genomics 250 (1996), S. 601-613 
    ISSN: 1617-4623
    Keywords: Sex-influenced modifier ; Position effect variegation ; Dosage effects ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Modifier of white (Mow), a dominant transacting gene, has been identified through a mutagenic screen for second-site loci that alter the level of expression of thewhite eye color locus.Mow reduces the expression ofwhite in most developmental stages, but enhances its expression in the pupal stage, the time at which the major contribution to the adult phenotype is made. Tests with anAlcohol dehydrogenase promoter-white reporter and a series ofwhite truncation constructs have shown thatMow fails to affect the reporter; cis-regulatory mutations ofwhite also show no response, suggesting a requirement forwhite regulatory domains for interaction withMow. A quantitative analysis of steady-state transcript levels reveals that thewhite mRNA level decreases in the presence of one dose ofMow in larvae and adults, but the reduction is greater in females than males. Two other functionally related genes,brown andscarlet, also exhibit a similar sexually dimorphic alteration in expression, mediated byMow. In the mid-pupal stage, by contrast, the level ofwhite andbrown mRNA is increased byMow. In addition,Mow acts as a weak suppressor of position effect variegation (PEV). These observations suggest a connection between dosage modulation of gene expression and suppression of position-effect variegation.
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  • 100
    ISSN: 1617-4623
    Keywords: Key words Aspartyl tRNA synthetase ; Tryptophanyl tRNA synthetase ; Drosophila melanogaster ; Sex-lethal ; Genetic modifiers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Stable activation of the Drosophila sex determination gene Sex-lethal in the female embryo is a multistep process. Early in embryogenesis Sex-lethal is regulated at the level of transcription, and then later in embryogenesis Sex-lethal regulation switches to an autoregulatory RNA splicing mechanism. Previous studies have shown that successful activation of Sxl requires both maternally and zygotically provided gene products, many of which are essential for viability and have other, non-sex specific functions. Using a screen for dosage-sensitive modifiers we identified a new maternally expressed gene, l(2)49Db, as a likely participant in Sxl activation. We show that the establishment of the Sxl autoregulatory splicing loop, but not the earlier steps in Sxl activation, is sensitive to the maternal dosage of l(2)49Db. We further demonstrate that l(2)49Db encodes an aspartyl tRNA synthetase. Finally we present evidence that this effect is indirect, by demonstrating that mutations in tryptophanyl tRNA synthetase are also dosage-sensitive maternal modifiers of Sex-lethal. These data suggest that stable activation of Sex-lethal in the embryo may be particularly sensitive to perturbation of the translational machinery.
    Type of Medium: Electronic Resource
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