Metabolic Studies with Banana Fruit Slices II. Effects of Inhibitors on Respiration, Ethylene Production, and Ripening
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Transverse slices of green banana fruit were vacuum� infiltrated with aqueous solutions of 24 potential inhibitors of protein synthesis, respiration, or ethylene production. The effects of these compounds were examined in the absence or presence of 10 p.p.m. ethylene. Of the compounds which produced marked effects mono� fiuoroacetate, 4�hydroxy�2�oxoglutarate (HKG), KCN, and cycloheximide were examined in more detail.Keywords:
Biological sciences
Puromycin
Dactinomycin
Mechanism of Action
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Cycloheximide is frequently presumed to inhibit specifically the cytoplasmic protein synthesis of eukaryotes. Although previous investigators have shown that it had other effects on the cells of a variety of organisms, these results were frequently presumed to be secondary effects of the inhibition of protein synthesis. This paper shows that a wide range of deleterious effects are produced by cycloheximide on a single organism, Chlamydomonas reinhardi Dangeard. If, protein synthesis is inhibited by nonpermissive conditions in temperature-sensitive mutants or with other treatments these "secondary" effects are not produced. Instead, cycloheximide appears to have two or three independent inhibitory effects on the cell. Moreover, in contrast to a number of previous investigations, these results show that protein synthesis is not required for RNA synthesis. Instead the rate of RNA synthesis is actually increased by interference with protein synthesis.
Chlamydomonas
Protein synthesis inhibitor
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Cycloheximide had no effect on multiplication of the meningopneumonitis agent in L cells in concentrations which eliminated over 90% of the protein synthesis in the host cells. Infected L cells treated with cycloheximide, however, incorporated labeled amino acids into the trichloroacetic acid-insoluble fraction. This incorporation was attributed to the biosynthetic activity of the meningopneumonitis agent. Synthesis of meningopneumonitis protein was abolished by chloramphenicol and chlortetracycline, inhibitors of bacterial protein synthesis, at concentrations which did not inhibit protein synthesis in L cells. Protein synthesis in the meningopneumonitis agent was sustained at a high rate when the host cells remained viable and declined as the L cells died. Overall host protein synthesis was not inhibited by multiplication of the meningopneumonitis agent.
Protein synthesis inhibitor
Trichloroacetic acid
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The respiratory behaviour of tomato fruits during their growth and ripening on the plant has been studied in relation to the season and light intensity during growth. Growth was found to be associated with an absolute increase in respiration rate. Fruits of all populations showed a marked rise and fall in respiration rate during ripening. This senescent drift was confirmed in detail by following the respiration of individual fruits ripening while still attached to the plant. Although exhibiting the usual senescent drift as they changed colour, fruits grown in summer cloth house plots showed a consistently lower rate of respiration than that observed in all other populations. It also has been shown that the chief path of gaseous exchange of the mature attached fruit is localized at the stem end.
Respiration rate
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Puromycin
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Abstract The effects of elevated temperatures upon protein biosynthesis were determined in L5178Y murine leukemic lymphoblasts. The rate of protein synthesis was inhibited proportionately to the increase in temperature. Efforts were made to determine the mechanism of heat inactivation of protein synthesis by studying the requirements for recovery of activity after the cells were returned to 37°C. The ability of actinomycin to block the recovery process suggests that elevated temperatures destroy or inactivate a species of RNA required for protein synthesis. Loss of RNA during heating of the cells is apparently at least partially dependent on protein synthesis, since the presence of cycloheximide during heat shock, is capable of ameliorating the effects of short duration heat treatment.
Lymphoblast
Protein synthesis inhibitor
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Streptomyces griseus
Protein synthesis inhibitor
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Protein synthesis inhibitor
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Based on evidence that 50% of herpes simplex 1 DNA is transcribed in HEp-2 cells in the absence of protein synthesis we examined the order and rates of synthesis of viral polypeptides in infected cells after reversal of cycloheximide- or puromycin-mediated inhibition of protein synthesis. These experiments showed that viral polypeptides formed three sequentially synthesized, coordinately regulated groups designated alpha, beta, and gamma. Specifically: (i) The alpha group, containing one minor structural and several nonstructural polypeptides, was synthesized at highest rates from 3 to 4 h postinfection in untreated cells and at diminishing rates thereafter. The beta group, also containing minor structural and nonstructural polypeptides, was synthesized at highest rates from 5 to 7 h and at decreasing rates thereafter. The gamma group containing major structural polypeptides was synthesized at increasing rates until at least 12 h postinfection. (ii) The synthesis of alpha polypeptides did not require prior infected cell protein synthesis. In contrast, the synthesis of beta polypeptides required both prior alpha polypeptide synthesis as well as new RNA synthesis, since the addition of actinomycin D immediately after removal of cycloheximide precluded beta polypeptide synthesis. The function supplied by the alpha polypeptides was stable since interruption of protein synthesis after alpha polypeptide synthesis began and before beta polypeptides were made did not prevent the immediate synthesis of beta polypeptides once the drug was withdrawn. The requirement of gamma polypeptide synthesis for prior synthesis of beta polypeptides seemed to be similar to that of beta polypeptides for prior synthesis of the alpha group. (iii) The rates of synthesis of alpha polypeptides were highest immediately after removal of cycloheximide and declined thereafter concomitant with the initiation of beta polypeptide synthesis; this decline in alpha polypeptide synthesis was less rapid in the presence of actinomycin D which prevented the appearance of beta and gamma polypeptides. The decrease in rates of synthesis of beta polypeptides normally occurring after 7 h postinfection was also less rapid in the presence of actinomycin D than in its absence, whereas ongoing synthesis of gamma polypeptides at this time was rapidly reduced by actinomycin D. (iv) Inhibitors of DNA synthesis (cytosine arabinoside or hydroxyurea) did not prevent the synthesis of alpha, beta, or gamma polypeptides, but did reduce the amounts of gamma polypeptides made.
Puromycin
De novo synthesis
Protein synthesis inhibitor
Viral protein
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Protein synthesis in vivo was studied at various times after the administration of sublethal doses of cycloheximide to rats. Cycloheximide caused an inhibition, followed by a dose-and time-dependent stimulation, of incorportation of labelled precursor into proteins of the liver and kidney. The stimulation of protein synthesis at 24h was not due to a change of precursor pool or the specific radioactivity of the precursor used. During the stimulatory period, leucine incorporation into various cellular protein fractions varied; incorporation into total nuclear protein was the most affected.
Protein synthesis inhibitor
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