Inhibition of protein synthesis in intact mammalian cells by arachidonic acid.
Optimal translation initiation in intact mammalian cells requires sequestered intracellular Ca2+. Arachidonic acid, which releases sequestered Ca2+ from cells and isolated organelles, was studied to assess its potential role in the regulation of protein synthesis via Ca2+ mobilization. Unsaturated fatty acids at microM concentrations inhibited protein synthesis in intact GH2 pituitary, C6 glial tumour and HeLa cells in a manner dependent on degree of unsaturation and cell number. Arachidonate was generally the most, and the fully saturated arachidic acid the least, potent of the fatty acids tested. At 2 x 10(6) GH3 cells/ml, amino incorporation into a broad spectrum of polypeptides was inhibited by 80-90% by 10-20 microM fatty acid. Inhibition was maximal at 4-8 min and was attenuated by 1-2 h and more pronounced at lower pH. Protein synthesis was maximally inhibited when arachidonate mobilized approx. 40% of cell-associated Ca2+. At lower concentrations (10 microM) arachidonate suppressed translational initiation, with the inhibition being reversed as extracellular Ca2+ concentrations were increased to supraphysiological values. At higher concentrations (20 microM) arachidonate inhibited peptide-chain elongation in a Ca(2+)-independent manner. Arachidonate also blocked elongation in reticulocyte lysates. The effects of arachidonate in intact cells were reversible with time via its metabolism or by washes containing BSA. Sufficient arachidonate appears to be synthesized during ischaemic stress to inhibit translation by either mechanism.
Rotman
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Department of Pharmacology, UMDNJ-Robert Wood Johnson Medical School, Piscataway 08854.
Brostrom
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Brostrom
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D000596:Amino Acids / Q000378:metabolism; D016718:Arachidonic Acid / Q000494:pharmacology*; D001345:Autoradiography; D002118:Calcium / Q000378:metabolism; D002413:Cations, Divalent; D004537:Eicosanoic Acids / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007930:Leucine / Q000378:metabolism; D008715:Methionine / Q000378:metabolism; D011500:Protein Synthesis Inhibitors; D014407:Tumor Cells, Cultured
D000596:Amino Acids; D002413:Cations, Divalent; D004537:Eicosanoic Acids; D011500:Protein Synthesis Inhibitors; D016718:Arachidonic Acid; D008715:Methionine; D007930:Leucine; D002118:Calcium