Inhibition of protein synthesis by antagonists of calmodulin in Ehrlich ascites tumor cells.
Several recent publications indicate that Ca2+ is required for protein synthesis in mammalian cells, including the Ehrlich ascites tumor cell. The present communication examines whether the effects of Ca2+ might be mediated through calmodulin or a related protein. Four calmodulin antagonists belonging to different chemical categories were used to provide evidence of calmodulin involvement. Three of the antagonists inhibited protein synthesis in intact cells; 50% inhibitory concentrations were 10 microM calmidazolium, 12 microM N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W7) and 17.5 microM trifluoperazine (TFP). Initiation was preferentially inhibited as indicated by an increase in the 80S monomers accompanied by a significant disaggregation of polyribosomes. All the antagonists also inhibited protein synthesis initiation in the cell-free protein-synthesizing system; 50% inhibitory concentrations for compound 48/80, calmidazolium, TFP, and W7 were 10 microM, 125 microM, 300 microM and 500 microM, respectively. A weak analogue of W7 inhibited only 20% at 1000 microM. Inhibition in the cell-free system was reversed by the addition of exogenous calmodulin in all four cases. The levels of 43S complexes were significantly elevated with all four antagonists, indicating a block in the utilization of 43S complexes. The similarity of the effects of four distinct classes of antagonists and their ready reversal by exogenous calmodulin leads us to suggest that there may be a role for calmodulin or a very similar calcium-binding protein in protein synthesis.
Kumar
R V
RV
Department of Biochemistry, University of Rochester, New York.
Panniers
R
R
Wolfman
A
A
Henshaw
E C
EC
D000818:Animals; D002147:Calmodulin / Q000037:antagonists & inhibitors*; D002286:Carcinoma, Ehrlich Tumor / Q000188:drug therapy* / Q000378:metabolism; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D007093:Imidazoles / Q000494:pharmacology; D008715:Methionine / Q000378:metabolism; D010442:Peptide Chain Initiation, Translational / Q000187:drug effects*; D011132:Polyribosomes / Q000187:drug effects / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000187:drug effects; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D012343:RNA, Transfer / Q000378:metabolism; D013449:Sulfonamides / Q000494:pharmacology; D013462:Sulfur Radioisotopes; D014268:Trifluoperazine / Q000494:pharmacology
D002147:Calmodulin; D007093:Imidazoles; D011500:Protein Synthesis Inhibitors; D013449:Sulfonamides; D013462:Sulfur Radioisotopes; D014268:Trifluoperazine; C031938:calmidazolium; C017967:W 7; D012343:RNA, Transfer; D008715:Methionine