The effect of changes in the respiratory metabolism upon genome activity in Drosophila

Leenders, H.J.; Berendes, H. D.

doi:10.1007/bf00284892

Cited by 85 publications

(29 citation statements)

References 12 publications

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“…include: recovery from prolonged anaerobiosis [e.g., 2 hr under nitrogen (13)], treatment of isolated glands with uncouplers of oxidative phosphorylation [e.g., 2,4-dinitrophenol (14,15)], and treatment of glands with inhibitors of electron transport [e.g., rotenone (4)]. …”

Section: Resultsmentioning

confidence: 99%

“…Identi- in air for 90 min, or (c) under nitrogen for 2 hr followed by 45 min in air, before their salivary glands were dissected and labeled. The glands were cultured as described in Materials and Methods and, after lipid extraction, the dried glands were dissolved by boiling for 1.5-2 min in 50 gl of buffer (0.1 M phosphate, pH 6.9, 1% sodium dodecyl sulfate, 4 M urea, and 1% 2-mercaptoethanol). Aliquots were loaded onto 10% acrylamide-sodium dodecyl sulfate gels according to MacGillivray et al (21) and run at 16 mA.…”

Section: Resultsmentioning

confidence: 99%

“…During late larval and "prepupal" development, at least, these changes are under hormonal control (1,2). Puffing activity can also be controlled by certain environmental treatments; in particular a brief exposure of salivary glands, either in the intact animal or in culture, to high temperature induces specific puffs (3)(4)(5).…”

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Parallel changes in puffing activity and patterns of protein synthesis in salivary glands of Drosophila.

Lewis¹,

Helmsing²,

Ashburner³

1975

Proc. Natl. Acad. Sci. U.S.A.

161

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The The pattern of puffing activity in the polytene chromosomes of Drosophila, and other Diptera, changes in a complex way during development. During late larval and "prepupal" development, at least, these changes are under hormonal control (1, 2). Puffing activity can also be controlled by certain environmental treatments; in particular a brief exposure of salivary glands, either in the intact animal or in culture, to high temperature induces specific puffs (3-5).We report the changes in salivary gland protein synthesis that result from temperature shock and the correlated changes in puffing activity. They will enable a formal proof of the hypothesis that puffs are active gene loci (6), a proof that demands the genetic mapping of a putative puff product to the puff itself and the demonstration that mutations which specifically result in the loss of the puffs also result in the absence of its putative product. Preparation of Samples. At the end of the labeling period the radioactive medium was removed with a fine-tipped glass pipette and the glands were flooded with medium containing a 100-fold excess of nonradioactive methionine. If necessary they were then stored at -70°to await further processing. After the wash in nonradioactive medium the glands were washed in cold 10% trichloroacetic acid and then transferred into small glass test tubes. Then they were washed with 10% cold trichloroacetic acid (20 min Control Experiments. The extraction procedure we used is essentially that of Tissieres et al. (9). We have varied the extraction method (for example by using boiling 10% trichloroacetic acid or prolonged chloroform:methanol extraction at 60°) without affecting the results. We have also studied the possible effects of sodium dodecyl sulfate on protease activity by treatment of samples with either phenyl methyl sulfonyl fluoride (10) or L-1-tosylamido-2-phenylethylchlo-3604

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Parallel changes in puffing activity and patterns of protein synthesis in salivary glands of Drosophila.

Lewis¹,

Helmsing²,

Ashburner³

1975

Proc. Natl. Acad. Sci. U.S.A.

161

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“…The induced proteins are termed heat shock polypeptides (hsps). A variety of chemical compounds also elicit the same response (2, 15,17,20,22), suggesting that the heat shock response is probably a reaction to metabolic stress rather than to temperature per se.Until recently, most data on the heat shock phenomenon were derived from studies with Drosophila melanogaster. All organisms studied thus far, however, respond in a similar fashion (34).…”

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confidence: 99%

70-Kilodalton heat shock polypeptides from rainbow trout: characterization of cDNA sequences.

Kothary¹,

Jones²,

Candido³

1984

Mol. Cell. Biol.

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RTG-2 cells, a line of fibroblasts from rainbow trout (Salmo gairdnerii), are induced to synthesize a distinct set of heat-shock polypeptides after exposure to elevated temperature or to low concentrations of sodium arsenite. We isolated and characterized two cDNA sequences, THS70.7 and THS70.14, encoding partial information for two distinct species of 70-kilodalton heat shock polypeptide (hsp70) from these cells. These sequences are identical at 73.3% of the nucleotide positions in their regions of overlap, and their degree of sequence conservation at the polypeptide level is 88.1%. The two derived trout hsp7O polypeptide sequences show extensive homology with derived amino acid sequences for hsp7O polypeptides from Drosophila melanogaster and Saccharomyces cerevisiae. Northern blot analysis of RNA from arsenite-induced RTG-2 cells, with the trout hsp7O cDNAs as probes, revealed the presence of three hsp7O mRNA species. Southern blot analysis of trout testis DNA cleaved with various restriction endonucleases revealed a small number of bands hybridizing to the hsp7O cDNAs, suggesting the existence of a small family of hsp70 genes in this species. Finally, trout hsp70 cDNA sequences cross-hybridized with restriction fragments in genomic DNA from HeLa cells, bovine liver, Caenorhabditis elegans, and D. melanogaster.The heat shock response provides an excellent system for the study of the processes which accompany rapid gene induction in eucaryotic cells. When organisms are subjected to a heat shock, transcription of most genes is suppressed, and the expression of a novel set of proteins is enhanced (for reviews, see references 2, 34, and 39). The induced proteins are termed heat shock polypeptides (hsps). A variety of chemical compounds also elicit the same response (2, 15,17,20,22), suggesting that the heat shock response is probably a reaction to metabolic stress rather than to temperature per se.Until recently, most data on the heat shock phenomenon were derived from studies with Drosophila melanogaster. All organisms studied thus far, however, respond in a similar fashion (34). The highly conserved nature of this response across a wide range of species implies a fundamental role for the hsps in the stressed organism.The major hsps of most organisms fall into three classes: the small hsps (15 to 30 kilodaltons), the hsp70-like (60 to 70 kilodaltons) polypeptides, and the hsp83-like (80 to 90 kilodaltons) polypeptides. These three classes of hsps are strongly conserved. Antibodies against the hsp70 and hsp89 from chicken embryo fibroblasts cross-react with their counterparts from a wide variety of organisms (16). Similarly, cross-hybridization of hsp70 genes from different organisms has been observed (10, 14,24,29,36). The small hsps from both D. melanogaster and Caenorhabditis elegans share extensive amino acid sequence homology with the mammalian a-crystallins (12, 33).The five hsp70 genes from D. melanogaster have been cloned and sequenced (11, 13,30,42). Very little sequence information about the hsp7o genes...

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“…Inhibition of proteins and RNA suppressed the stimulatory effects of isocitrate and tyrosine, suggesting that some of the puffs in the polytene chromosomes reflected gene activity for the use of isocitrate and tyrosine for respiration during conditions of stress. 19 The sensitivity of our apparatus suggests that we might be able to assess O 2 consumption in salivary glands of individual mosquitoes uninfected or infected with Plasmodium sporozoites.…”

Section: Discussionmentioning

confidence: 99%

Automated measurement of oxygen consumption by the yellow fever mosquito, Aedes aegypti.

Diarra¹,

Roberts²,

Christensen³

1999

The American Journal of Tropical Medicine and Hygiene

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Abstract. Oxygen consumption of single mosquitoes was measured using a differential pressure transducer (DPT) connected to two small chambers. A mosquito was placed in the experimental chamber (P1) and was separated from NaOH by 4 cm 2 of marquisette mesh. The reference chamber (P2) contained the same amount of NaOH and the marquisette mesh but without a mosquito. When these two chambers were sealed, removed O 2 from P1 resulted in a pressure decrease with respect to P2. This pressure differential was transduced into an output voltage that was directly proportional to the amount of O 2 consumed by the mosquito. An array of eight DPTs was interfaced with an IBM 486 PC using an ADAC 5500MF analog to digital converter and software from ADAC (Direct View) to automate the recording procedure. We determined that our apparatus was sensitive enough to detect subtle differences in O 2 consumption in mosquitoes subjected to different physiologic conditions. Manometric methods have been used extensively in the study of respiration and have produced a great variety of apparatus to carry out these measurements. Among the many different respirometers that have been produced, the Warburg respirometer has been the most widely used.1 In the early 1960s, Gilson 2 constructed a simpler differential respirometer with a single reference flask, for multiple active flasks, and a digital readout volumometer to circumvent the tedious calculation and calibration steps. The use of manometric methods, however, has decreased because of their complexity, fragility, and cumbersome nature 2 , and were replaced by the more sensitive, accurate, simple, and rapid spectrometric methods. Spectrometric methods, although well suited to the study of chemical and biological reactions at the cellular level, are not well suited for studies of whole organisms.In an attempt to understand why energy costs do not balance in insects, Wightman 3 traced the inaccuracy in the R term (respiration). He found that R was always lower when measured with a respirometer than the value obtained from consumption minus production and egesta. This suggested that of all the parameters of an energy budget, respiration was the most difficult to obtain because of the numerous factors affecting it in unpredictable ways. This was even more true for social insects. Although respiration of ant colonies did not suggest any circadian rhythm, it was possible, to observe an annual respiratory pattern characterized by a lower oxygen consumption during the cold seasons.4, 5 Termites had a more complex respiratory system in which both the organism and its bacterial endosymbionts had to be considered. In the latter system, measurements of oxygen consumption have been used to study the possible roles of glucose, xylose, pyruvate, alanine, ␣-ketoglutarate, and acetate as energy sources for both the termite and its endosymbiotic bacterium. 6 Metabolic studies of non-social insects, such as the house cricket, showed a strong correlation between mating and the total amount of oxygen consumed b...

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The effect of changes in the respiratory metabolism upon genome activity in Drosophila

Cited by 85 publications

References 12 publications

Parallel changes in puffing activity and patterns of protein synthesis in salivary glands of Drosophila.

Parallel changes in puffing activity and patterns of protein synthesis in salivary glands of Drosophila.

70-Kilodalton heat shock polypeptides from rainbow trout: characterization of cDNA sequences.

Automated measurement of oxygen consumption by the yellow fever mosquito, Aedes aegypti.

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