Macromolecular synthesis was studied in individual liver cells rendered permeable to macromolecules and charged molecules by treatment with toluene. Toluene-treated cells were compared to intact cells with regard to their ability to synthesize protein, RNA, and DNA. The permeable cells catalyzed the incorporation of amino acids into protein in a system which was sensitive to cycloheximide. Maximal incorporation required the addition of tRNA, ATP, GTP, an energy source and various cations. RNA synthesis also took place in these cells and was inhibited by actinomycin D. Maximal incorporation required all four ribonucleoside triphosphates, an energy-generating system, and Mn2+, K+, and F-. The toluene-treated cells also were active for DNA synthesis when Ca2+ was present to induce endonucleolytic cleavage of the endogenous DNA. For maximal synthesis, all four deoxyribonucleoside triphosphates, ATP, K+, Mg2+, polyamines, and mercaptoethanol were required. These studies serve to emphasize the potential usefulness of toluene treatment for studying biosynthetic processes in mammalian cells.
ASJC Scopus subject areas
- Molecular Biology