Polypeptide Synthesis in Toluene‐Treated Lymphocytes

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 tha...

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Autor principal: BURRONE, O.R.
Formato: JOUR
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Acceso en línea:http://hdl.handle.net/20.500.12110/paper_00142956_v86_n2_p439_BURRONE
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Sumario: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 10000 ×g extracts. Such a process occurs inside the treated cells as demonstrated by the fact that [14C]uridine‐labelled ribosomes remain associated with the toluene‐treated lymphocytes even after incubation at 37 °C. A nuclease from Staphylococcus aureus was able to penetrate the permeabilized cells and to break the polysome‐bound endogenous messenger RNA. However, the protein‐synthesizing machinery inside the toluene‐treated lymphocytes was unaffected by the nuclease, as demonstrated by the unimpairement of polyphenylalanine synthesis when poly(U) was added after the preincubation with the enzyme. These results suggest that the toluene treatment can be considered as an important tool for the study of the synthesis of macromolecules and its regulation in eukaryotic cells. Copyright © 1978, Wiley Blackwell. All rights reserved