Evidence for rapid limitation of the I element copy number in a genome submitted to several generations of I-R hybrid dysgenesis in Drosophila melanogaster

Summary When Drosophila melanogaster males coming from a class of strains known as inducer are crossed with females from the complementary class (reactive), a quite specific kind of sterility is observed in the F₁ female progeny (denoted SF). The inducer chromosomes differ from the reactive chromosomes by the presence of a transposable element (called the I factor) that is responsible for the induction of this dysgenic symptom. In the germ line of dysgenic females, up to 100% of the reactive chromosomes may be contaminated, i.e. they acquire I factor(s) owing to very frequent replicative transpositions. A contaminated reactive stock was obtained by reconstructing the reactive genotype in the offspring of SF females and its kinetics of invasion by I elements was followed in the successive inbred dysgenic generations. The results show that the mean copy number of I elements increased very quickly up to the level of inducer strains and then stayed in equilibrium even though the dysgenic state was perpetuated by selection for SF sterility at every generation. The possible mechanisms of this copy number limitation are discussed.     

Formation and activity of covalent conjugates of poliovirus and ligands binding to cell surface structures

Disulfide-linked conjugates of poliovirus with streptavidin or concanavalin A were formed and the binding of the conjugates to mouse L cells that lack natural poliovirus receptors was studied. The conjugate with streptavidin was specifically bound to biotinylated L cells, but not to unmodified L cells. The conjugate with conA was bound to L cells in the absence of, but not in the presence of alpha-methyl mannoside. Incubation of L cells with bound conjugates did not produce virus, although the conjugates were highly infectious in HeLa cells, containing natural poliovirus receptors. This suggests that the artificially bound virus was unable to penetrate the L cells and start replication. The possibility that binding of the virus to the natural receptor is required for efficient infection is discussed.     

Separation of multiple components of HeLa cell nuclear extracts required for pre-messenger RNA splicing

Components essential for nuclear pre-messenger RNA splicing have been partially purified from HeLa cell nuclear extracts by chromatography on DEAE-Sepharose, heparin-Sepharose, Mono Q, and Mono S. We have obtained six fractions which, when combined, efficiently splice a synthetic adenovirus 2 major late RNA substrate in vitro. All fractions contain components that support the formation of splicing intermediates (the cleaved 5' exon and the intron-exon 2 lariat). At least one of the fractions also contains an activity that is essential for the second step in the splicing reaction, namely cleavage at the 3' splice site and exon ligation. Two of the fractions are enriched in the major small nuclear ribonucleoprotein particles U1, U2, U4/U6, and U5. They participate in the formation of the splicing complexes which precedes the cleavage and ligation reactions. The remaining four fractions appear to contain protein factors, as suggested by their resistance to micrococcal nuclease.     

Actinomycin synthetases. Multifunctional enzymes responsible for the synthesis of the peptide chains of actinomycin

Two enzymes were purified from actinomycin-synthesizing Streptomyces chrysomallus which could be identified as peptide synthetases involved in the biosynthesis of actinomycin. Actinomycin synthetase II activates the first two amino acids of the peptide chains of the peptide lactone antibiotic, threonine and valine (or isoleucine), as thioesters via their corresponding adenylates. It is a single polypeptide chain of Mr 225,000. Similarly, actinomycin synthetase III activates proline, glycine, and valine (the remaining three amino acids in the antibiotic) as thioesters and is a single polypeptide chain of about Mr 280,000. It also carries the methyltransferase function(s) for N-methylation of thioesterified glycine and valine. In addition, it catalyzes the formation of cyclo(sarcosyl-N-methyl-L-valine) from glycine, L-valine, and S-adenosyl-L-methionine at the expense of ATP. Although the cell-free synthesis of the peptide lactone was not as yet accomplished, the data provide evidence that together with the 4-methyl-3-hydroxyanthranilic acid-activating enzyme (now designated as actinomycin synthetase I) all amino acid-activating protein components of the actinomycin-synthesizing enzyme complex are identified.     

Modulating effect of cystamine and unsaturated fatty acids on gamma-glutamyl transpeptidase: Relation to cellular oxidative status

Summary Cell surface gamma-glutamyl transpeptidese activity in cultured neoplastic astrocytes was significantly increased upon treatment of the cells with the hepatoprotective disulfide, cystamine. The cystamine effect was sensitive to cycloheximide and could be significantly depressed by exogenous glutathione. Surface gamma-glutamyl transpeptidase activity was also modulated by the presence in the culture medium of the unsaturated fatty acids, linoleic acid and arachidonic acid. Metabolism of the fatty acids via the cyclooxygenase pathway was not a prerequisite for their modulation of the glycoprotein ectoenzyme. Lipoxygenase, however, was found to potentiate the unsaturated fatty acid effect in neoplastic astrocytes. Lipoxygenase is reported to catalyze the conversion of unsaturated fatty acids to their corresponding peroxides. The data indicate an oxidative influence on the control of gamma-glutamyl transpeptidase activity.     

Axonal Transport Characteristics of Gangliosides in Sensory Axons of Rat Sciatic Nerve

The distribution of axonally transported gangliosides and glycoproteins along the sciatic nerve was examined from 3 h to 4 weeks following injection of[3H]glucosamine into the fifth lumbar dorsal root ganglion of adult rats. Incorporation of labeled precursor into these glycoconjugates reached a maximal level in the ganglion within 6 h. Outflow patterns of radioactivity for glycoproteins showed a well-defined crest with a transport rate of approximately 330 mm/day. In contrast, the crest of transported gangliosides was continuously attenuated, implying a significant deposition along the axon, and an alternative method of calculating velocity was required. Analysis of accumulation of labeled material at double ligatures demonstrated both anterograde and retrograde transport of glycoproteins and gangliosides and allowed for the calculation of an anterograde transport rate of about 270 mm/day for each. Additional evidence of ganglioside transport is provided in that the TLC pattern of transported radioactive gangliosides accumulating at a ligature is significantly different from the pattern seen in the dorsal root ganglion or following intraneural administration of the labeled precursor. These data indicate that gangliosides are transported at the same rapid rate as glycoproteins but are subject to a more extensive exchange with stationary material than are glycoproteins.     

Enzymatic Protection Against Peroxidative Damage in Isolated Brain Capillaries

The content of polyunsaturated fatty acids, the activities of superoxide dismutase (SOD), glutathione peroxidase, glutathione reductase, and catalase, and the concentration of reduced glutathione were measured in cerebral microvessels isolated from rat brain. Polyunsaturated fatty acids, mainly arachidonic, linoleic, and docosahexaenoic acids, accounted for 32% of total fatty acids in cerebral microvessels. Whereas total SOD activity in the microvessels was slightly lower than that found in cerebrum and cerebellum, glutathione peroxidase and glutathione reductase activities were twice as high and catalase activity was four times higher. Glutathione peroxidase in microvessels is active on both hydrogen peroxide and cumen hydroperoxide, and it is strongly inhibited by mercaptosuccinate. After several hours of preparation, the concentration of reduced glutathione in isolated microvessels was 0.7 mumol/mg of protein, which corresponds to a concentration of approximately 3.5 mM. Our results indicate that the blood-brain barrier contains large amounts of peroxide-detoxifying enzymes, which may act, in vivo, to protect its highly polyunsaturated membranes against oxidative alterations.     

Binding of anti-fibronectin to early amphibian ectoderm does not result in inhibition of neural induction under in vitro conditions

Summary Antibodies directed to fibronectin (anti-FN) were injected into the blastocoel of late blastulae of Xenopus laevis. Two animal caps (ectoderm) were isolated, when control embryos reached the early gastrula stage, and were combined with untreated upper blastopore lip in the sandwich method. In two control series fibronectin or Holtfreter solution was injected into the blastocoel. The results of the experiments suggest that neural induction cannot be prevented by binding anti-FN to fibronectin, which covers the blastocoelic side of the ectoderm. The data support the view that extracellular matrix proteins are not themselves responsible for neural induction. However, in comparison with the control series a slight shift of the differentiation pattern in the spinocaudal direction could be observed in the anti-FN series. The possible role of extracellular proteins in the formation of a close juxtaposition of mesodermal and ectodermal target cells as a prerequisite for shortdistance transmission of neural inducers is discussed.     

Influence of leaf surfaces on movements by the hymenopterous parasitoid Trichogramma exiguum

Walking speeds of female Trichogramma exiguum Pinto & Platner were fastest on maize and soybean (12 cm/min), intermediate on tomato (8 cm/min), and slowest on woolly mullein, Verbascum thapsus (3 cm/min). Similarly, rates of turning along the paths of walking T. exiguum were smallest on maize (median angle=0°±15°), intermediate on soybean and tomato, and greatest on V. thapsus (median angle=30°±15°). Leaf trichome density and morphology influenced walking behavior. Walking was slowed and flight initiation delayed for T. exiguum walking on Amaranthus hybridus leaves compared to either maize or filter paper. When inundative releases are conducted, the effects of plant surfaces on searching rates and arrestment should be considered in determining release rates of Trichogramma spp.

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Résumé La vitesse de marche a été déterminée en lâchant des individus sur chaque substrat végétal et en traçant leur parcours sur une plaque de verre placée à 9 mm au dessus du sujet. Les déviations angulaires de portions successives des tracés, longues de 1 mm, ont été utilisées pour mesurer les taux de changement de direction. Dans une deuxième expérience, des individus ont été lâchés au milieu d'une cercle de 40 mm de diamètre sur chaque substrat et les temps écoulés avant l'envol ou pour atteindre le bord du cercle à la marche ont servi à évaluer respectivement la propension au vol et la vitesse de déambulation.Les vitesses de marche ont été plus rapides sur maïs et soja (12 cm/min), moyennes sur tomate (8 cm/min) et les plus lentes sur Verbascum thapsus (Scrophulariaceae) (3 cm/min). De le même façon, les angles de changement de direction au cours des marches effectuées par T. exiguum ont été plus petits sur maïs, moyens sur soja et tomate, et plus grands sur V. thapsus. Chez T. exiguum marchant sur des feuilles d'Amaranthus hybridus L. (Amaranthaceae), la marche a été plus lente et l'envol plus tardif que sur maîs ou papier filtre.Lors de lâchers inondatifs, les effets des surfaces végétales sur les vitesses d'exploration et d'arrêt devraient être pris en compte pour déterminer les vitesses de lâcher des Trichogramma spp.