The naturally occurring polyamines spermine and spermidine stimulate in low concentrations the amino acid-incorporating activity of cell-free systems from cerebral cortex, cerebellum and liver from the rat to a significant degree, but in high concentrations they have strong inhibiting effects on these systems. Maximal polyamine stimulation is observed with suboptimal Mg2+ concentrations in the reaction mixtures; increasing Mg2+ to 15 mM results in an inhibition of cell-free protein synthesis by polyamines even at low concentrations. The stimulating effect of spermine and spermidine involves a facilitated binding of template RNA and aminoacylated tRNA to the ribosomes. In contrast, the aminoacylation of tRNA is not influenced. These data indicate that spermine as well as spermidine have regulatory functions during the translational process in cerebral cortex, cerebellum and liver. They seem to interact with ribosomal functions and to cause a shift in the optimal Mg2+ concentration required for the synthesis of brain and liver proteins.