Polypeptide Synthesis in Toluene‐Treated Lymphocytes

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Abstract
Normal human lymphocytes stimulated by phytohemagglutinin P, and other mammalian cells were rendered permeable to macromolecules such as poly(U) and proteins, by treatment with a low concentration of toluene. Under this condition, poly(U) translation was more efficient in the permeabilized cells than in 10,000 .times. g extracts. Such a process occurred inside the treated cells as demonstrated by the [14C]uridine-labeled ribosomes that remained associated with the toluene-treated lymphocytes even after incubation at 37.degree. C. A nuclease from Staphylococcus aureus was able to penetrate the permeabilized cells and to break the polysome-bound endogenous mRNA. The protein-synthesizing machinery inside the toluene-treated lymphocytes was unaffected by the nuclease, as demonstrated by the unimpairment of polyphenylalanine synthesis when poly(U) was added after the preincubation with the enzyme. The toluene treatment can be considered as an important tool for the study of the synthesis of macromolecules and its regulation in eukaryotic cells.