DNA synthesis in detergent-treated mouse ascites sarcoma cells.
Mouse ascites sarcoma cells (SR-C3H/He cells) were made  permeable to nucleoside triphosphates by treatment with nonionic detergents in a nearly isotonic condition. The permeable cells synthesized DNA in the presence of the four deoxyribonucleoside triphosphates, ATP, Mg2+, and the proper ionic environment. The optimum detergent concentration for DNA synthesis was 0.015--0.020% with Triton X-100, 0.020% with Nonidet P-40, and about 0.0025% with Brij 58. Higher concentrations of detergents were rather inhibitory to DNA synthesis. DNA synthesis in Triton-permeabilized cells was thought to be replicative, and the activity in the optimum conditions was much higher than that measured in hypotonic permeable cells or in isolated nuclei. These studies show the potential usefulness of detergent treatment for examining DNA replication in mammalian cells in vitro.
D000818:Animals; D002478:Cells, Cultured; D004261:DNA Replication; D004273:DNA, Neoplasm / Q000096:biosynthesis*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology*; D007038:Hypotonic Solutions; D051379:Mice; D008809:Mice, Inbred C3H; D011092:Polyethylene Glycols / Q000494:pharmacology; D001357:Sarcoma, Avian / Q000378:metabolism*; D013942:Thymine Nucleotides / Q000378:metabolism
D004273:DNA, Neoplasm; D003902:Detergents; D007038:Hypotonic Solutions; D013942:Thymine Nucleotides; D011092:Polyethylene Glycols; D004259:DNA-Directed DNA Polymerase