Replication and Repair of DNA in Cells of Escherichia coli Treated with Toluene

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Abstract
DNA synthesis has been studied in Escherichia coli cells made permeable to nucleotides by treatment with toluene. Replicative synthesis, as distinguished from repair synthesis, occurs at a rate comparable to that observed in vivo; it is dependent on the presence of all four deoxyribonucleoside triphosphates, but does not require exogenous DNA; and it is stimulated by ATP. Furthermore, replicative synthesis can be abolished at the restrictive temperature in DNA temperature-sensitive mutants. N-ethylmaleimide completely inhibits this type of synthesis, whereas it does not inhibit repair synthesis. Repair synthesis further differs from replicative synthesis in the following points: it does not require ATP; it persists at the restrictive temperature in DNA temperature-sensitive mutants; it can be induced by endogenous or exogenous nuclease activity; and its demonstration requires a Pol(+) strain.