Isolation of twelve satellite DNAs from Drosophila hydei

The genome of a Drosophila hydei genotype with a reduced amount of heterochromatin was fractionated by three cycles of preparative gradients: firstly in Ag⁺/Cs₂SO₄, secondly in actimomycin D/CsCl, and finally in neutral CsCl. Using this method, twelve highly repetitive simple-sequence satellites were isolated. Ten of them comprised only a minor amount of the genome in contrast to the two major satellites found earlier¹ (p = 1.696 and 1,714 g/cm³). These minor satellites were characterized by their banding in the gradient systems used, by their density in neutral CsCl, and by their melting point. Using these characteristics, it was found that the fractions of the Ag^?/Cs₂SO₄ gradient do not contain purified single components, because up to five different satellites band in the same position of the Ag^?/Cs₂SO₄ gradient. It was possible to isolate a high number of satellites even from a genome with a reduced amount of heterochromatin. Thus, the D hydei heterochromatin does not domain one unique highly repetitive sequence DNA, but is comprised of many different satellite sequences.     

A mating plug protein reduces early female remating in Drosophila melanogaster

Mating plugs are formed within the female reproductive tract during mating from male ejaculate constituents or even from male genitalia themselves. Across species, mating plugs have roles in sperm storage and the prevention of female remating. In the fruitfly Drosophila melanogaster, accessory gland proteins such as the sex peptide are known to reduce female remating, however this effect can take some time to establish, hence other ejaculate components must also be involved. We hypothesised a role for the PEBII mating plug protein in the prevention of early female remating. Using RNA interference we produced PEBII knockdown males. We found that these males were significantly less able to prevent female remating in the 4 h following mating. The mating plugs produced by PEBII knockdown males also showed lower levels of autofluorescence in the first 10 min after the start of mating, suggesting they differed in composition to those of control males. Reduced levels of PEBII had no effect, however, on fecundity, progeny production or egg-adult viability in the first 24 after mating, suggesting there were no short-term effects of PEB II on sperm transfer, storage or use. Our results show that PEBII has a subtle but significant role in the prevention of early female remating.     

The behavioural basis of overdominance in competitive mating success at the ebony locus of Drosophila melanogaster

Male files homozygous for the gene ebony¹¹ are partially blind, and at a disadvantage in competitive mating. The courtship of the mutant males is deficient in wing vibration stimulation, which is characterized by a low proportion of sine song and a high intra-pulse frequency. Males heterozygous for ebony have normal vision, but show an increase in courtship song, and are superior in competitive mating to wild type males. The auditory characteristics of courtship song produced by heterozygous males are indistinguishable from those of wild type, and their superiority in competitive mating success is due to overdominance involving this specific element of male courtship behaviour.     

Male Drosophila melanogaster show adaptive mating bias in response to female infection status

Given the non-trivial cost of reproduction for males and substantial variation in female quality, males have been predicted to show mating bias as an evolved strategy. Using a large outbred population of Drosophila melanogaster, we test this prediction and show that males may adaptively bias their mating effort in response to the infection status of females. Given a simultaneous choice between females infected with pathogenic bacteria and sham infected females, males preferentially mated with the latter, who had a higher reproductive output compared to infected females. This may provide evidence for pre-copulatory male mate choice. Assessment of the reproductive behaviour ensured that the observed pattern of mating bias was not due to differences in receptivity between females infected with pathogenic bacteria and sham infected females. Further, there was no evidence for post-copulatory male mate choice measured in terms of copulation duration.     

Cyclical expression of Na/K-ATPase in the visual system of Drosophila melanogaster

In the first (lamina) and second (medulla) optic neuropils of Drosophila melanogaster, sodium pump subunit expression changes during the day and night, controlled by a circadian clock. We examined α-subunit expression from the intensity of immunolabeling. For the β-subunit, encoded by Nervana 2 (Nrv2), we used Nrv2-GAL4 to drive expression of GFP, and measured the resultant fluorescence in whole heads and specific optic lobe cells. All optic neuropils express the α-subunit, highest at the beginning of night in both lamina and medulla in day/night condition and the oscillation was maintained in constant darkness. This rhythm was lacking in the clock arrhythmic per⁰ mutant. GFP driven by Nrv2 was mostly detected in glial cells, mainly in the medulla. There, GFP expression occurs in medulla neuropil glia (MNGl), which express the clock gene per, and which closely contact the terminals of clock neurons immunoreactive to pigment dispersing factor. GFP fluorescence exhibited circadian oscillation in whole heads from Nrv2-GAL4 + UAS-S65T-GFP flies, although significant GFP oscillations were lacking in MNGl, as they were for both subunit mRNAs in whole-head homogenates. In the dissected brain tissues, however, the mRNA of the α-subunit showed a robust daily rhythm in concentration changes while changes in the β-subunit mRNA were weaker and not statistically significant. Thus in the brain, the genes for the sodium pump subunits, at least the one encoding the α-subunit, seem to be clock-controlled and the abundance of their corresponding proteins mirrors daily changes in mRNA, showing cyclical accumulation in cells.     

Multiple function of pteridines in Drosophila: The fluorescence of the ejaculatory bulb in Drosophila melanogaster

It is demonstrated that the strong fluorescence of the ejaculatory bulb of Drosophila melanogaster males is caused by the presence of pteridines. The pteridine composition in the bulb is affected by the mutations ry² and ma-l^F1 in which isoxanthopterin has also been detected. Our results show that the bulbs of wild-type and white-eyed mutant males possess the same pteridines. Some data suggest that the bulbal pteridines originate from the testis region. Partly on the basis of former histochemical findings it is suggested that in the bulbal cavity the pH is high favouring the fluorescent dihydro-states of the pteridines present. All these and additional literature data on the ejaculatory bulb are discussed in connection with various biological processes. Some internal larval structures in which pteridines play or might play a functional role were found to present autofluorescence.     

Characterization of the Drosophila melanogaster Dis3 ribonuclease

The Dis3 ribonuclease is a member of the hydrolytic RNR protein family. Although much progress has been made in understanding the structure, function, and enzymatic activities of prokaryotic RNR family members RNase II and RNase R, there are no activity studies of the metazoan ortholog, Dis3. Here, we characterize the activity of the Drosophila melanogaster Dis3 (dDis3) protein. We find that dDis3 is active in the presence of various monovalent and divalent cations, and requires divalent cations for activity. dDis3 hydrolyzes compositionally distinct RNA substrates, yet releases different products depending upon the substrate. Additionally, dDis3 remains active when lacking N-terminal domains, suggesting that an independent active site resides in the C-terminus of the protein. Finally, a study of dDis3 interactions with dRrp6 and core exosome subunits in extracts revealed sensitivity to higher divalent cation concentrations and detergent, suggesting the presence of both ionic and hydrophobic interactions in dDis3-exosome complexes. Our study thus broadens our mechanistic understanding of the general ribonuclease activity of Dis3 and RNR family members.     

MMS sensitive strains in Drosophila melanogaster

Larval populations of Drosophila melanogaster were treated with the monofunctional alkylating agent methyl methyl methanesulfonate (MMS). Four different stocks were tested. The method methyl used permits the direct registration of MMS-induced lethality of every genotype present in the treated population. Up to 5-fold differences in sensitivity between different genotypes were obeserved. Larvae of the wild-type strain. Antibes, which was reported to have increased UV sensitivity in embryonic stages, were MMS sensitive. In another stock, marked with spa^pol, female larvae were about twice as sensitive as male larvae.     

Drosophila melanogaster deoxyribonucleoside kinase activates gemcitabine

Drosophila melanogaster multisubstrate deoxyribonucleoside kinase (Dm-dNK) can additionally sensitize human cancer cell lines towards the anti-cancer drug gemcitabine. We show that this property is based on the Dm-dNK ability to efficiently phosphorylate gemcitabine. The 2.2 Å resolution structure of Dm-dNK in complex with gemcitabine shows that the residues Tyr70 and Arg105 play a crucial role in the firm positioning of gemcitabine by extra interactions made by the fluoride atoms. This explains why gemcitabine is a good substrate for Dm-dNK.     

Drosopterins in phototaxis-selected strains of Drosophila melanogaster

The age-dependent drosopterin concentration was investigated in phototaxis-selected strains (Röko, Röpo, Röne, Stab) of D. melanogaster using extracted head homogenates. The results are compared to the wild stock derived strain Plus. The drosopterin concentration increases with the age of the flies. Females have larger concentrations than males. There are no differences in the drosopterin concentrations among the phototaxis-selected strains. Plus shows a higher drosopterin content than all other strains. This difference might be based on the genetical background, which is different in these strains. The data indicate that the phototactic behaviour is not related to the drosopterin concentration of the eyes.     

Mate recognition in members of the Drosophila nasuta complex

An examination of the courtship patterns of species belonging to the Drosophila nasuta complex shows that markings which are characteristic of males are almost certainly involved in their recognition as conspecifics by females. In each species the males perform a courtship sequence in such a way as to expose these markings to the female. A phylogeny of the species based on both male courtship patterns and the associated male markings is constructed. This phylogeny is compared to previously published phylogenies of the same group which were based on fixed and floating chromosomal differences between the species. Finally, the evidence which the complex provides for the utility and general significance of a new species concept, the Recognition Concept, is evaluated.     

Mutagenicity tests with metrifonate in Drosophila melanogaster

The organophosphorus insecticide metrifonate (O,O-dimethyl(1-hydroxy-2,2,2-trichloroethyl)phosphonate), also known as trichlorfon or Dipterex ®, was tested for its ability to induce sex-linked and autosomal recessive lethal mutations in Drosophila melanogaster. There was no evidence of an increase in the frequency of lethal mutations after feeding adult males with metrifonate, but the high toxicity of the compound meant that only low concentrations could be used in this test system.     

Molecular characterization of choline acetyltransferase from Drosophila melanogaster

Purified acetyl-CoA: choline O-acetyltransferase (EC 2.3.1.6) from Drosophila melanogaster has been shown to contain two major polypeptides of 67 and 54 K Daltons. However, all enzyme activity is found in a single molecular weight form of approx 67 K Daltons as determined by sucrose gradient sedimentation and molecular exclusion chromatography. The latter showed both the 67 and 54 K Dalton polypeptides on polyacrylamide gel electrophoresis in sodium lauryl sulfate (10% acrylamide). Analysis of purified choline acetyltransferase on polyacrylamide gel electrophoresis in sodium lauryl sulfate (15% acrylamide) revealed the presence of an additional polypeptide at 13 K Daltons. Tryptic-peptide maps of the 67, 54 and 13 K Dalton components showed all three to be structurally related. In addition to several common tryptic peptides, the 13 K Dalton polypeptide contained three tryptic-peptides that were also found in the 67 K Dalton polypeptide, but were absent from the 54 K Dalton polypeptide. This evidence suggests that native Drosophila choline acetyltransferase may exist in two forms, one a single polypeptide chain with a molecular weight of 67 K Daltons and the other consisting of two noncovalently bound polypeptide chains with molecular weights of 54 and 13 K Daltons. The latter form is the major one isolated and may be generated by limited proteolysis of the single chain 67 K Dalton form.