Translational suppression by Ca2+ ionophores : Reversibility and roles of Ca2+ mobilization, Ca2+ influx, and nucleotide depletion

• Published in: Cell biology and toxicology 1996-04, Vol.12 (2), p.101-113
• Main Authors: GMITTER, D, BROSTROM, C. O, BROSTROM, M. A
• Format: Article
• Language: English
• Subjects: Adenosine Triphosphate - metabolism; Animals; Biological and medical sciences; Blood Proteins - pharmacology; Calcimycin - pharmacology; Calcium - metabolism; Detergents - pharmacology; Drug toxicity and drugs side effects treatment; Guanosine Triphosphate - metabolism; Ionophores - pharmacology; Medical sciences; Mice; Miscellaneous (drug allergy, mutagens, teratogens...); Mitochondria - drug effects; Mitochondria - metabolism; Nucleotides - deficiency; Nucleotides - metabolism; Pharmacology. Drug treatments; Pituitary Neoplasms; Polyethylene Glycols - pharmacology; Protein Biosynthesis - drug effects; Sclerosing Solutions - pharmacology; Serum Albumin - pharmacology; Tumor Cells, Cultured - drug effects; Tumor Cells, Cultured - metabolism
• ISSN: 0742-2091; 1573-6822
• DOI: 10.1007/bf00143360

The divalent cation selective ionophores A23187 and ionomycin were compared for their effects on the Ca2+ contents, nucleotide contents, and protein synthetic rates of several types of cultured cells. Both ionophores reduced amino acid incorporation by approximately 85% at low concentrations (50-300 nmol/L) in cultured mammalian cells without reducing ATP or GTP contents. At these concentrations A23187 and ionomycin each promoted substantial Ca2+ efflux, whereas at higher concentrations a large influx of the cation was observed. Ca2+ influx occurred at lower ionophore concentrations and to greater extents in C6 glioma and P3X63Ag8 myeloma than in GH3 pituitary cells. The ATP and GTP contents of the cells and their ability to adhere to growth surfaces declined sharply at ionophore concentrations producing increased Ca2+ influx. Prominent reductions of nucleotide contents occurred in EGTA-containing media that were further accentuated by extracellular Ca2+. Ionomycin produced more Ca2+ influx and nucleotide decline than comparable concentrations of A23187. The inhibition of amino acid incorporation and mobilization of cell-associated Ca2+ by ionomycin were readily reversed in GH3 cells by fatty acid-free bovine serum albumin, whereas the effects of A23187 were only partially reversed. Amino acid incorporation was further suppressed by ionophore concentrations depleting nucleotide contents. Mitochondrial uncouplers potentiated Ca2+ accumulation in response to both ionophores. At cytotoxic concentrations Lubrol PX abolished protein synthesis but did not cause Ca2+ influx. Nucleotide depletion at high ionophore concentrations is proposed to result from increased plasmalemmal Ca2+-ATPase activity and dissipation of mitochondrial proton gradients and to cause intracellular Ca2+ accumulation. Increased Ca2+ contents in response to Ca2+ ionophores are proposed as an indicator of ionophore-induced cytotoxicity.