[Location of RNA-dependent DNA polymerase within the endogenous template complex and discrimination of RNA- and DNA-dependent synthesis of DNA in Escherichia coli cells by alkylating dTTP gamma-amide].

These data demonstrate that actinomycin D inhibits only 75-80% of DNA synthesis in cells of E. coli treated by lysozyme and ethyl enediaminetetraacetate. The residual synthesis is not the result of untemplated polymerization of dNTP. The DNA synthesis in spheroplasts does not correlate with replication of chromosomal DNA of E. coli catalyzed by DNA polymerase III sensitive to sulfhydryl blocking agents. N-ethylmaleimide does not inhibit this synthesis. No ATP stimulation of DNA synthesis is observed. The enzyme(s) responsible for DNA synthesis on endogenous template is (are) concentrated in interphase (D-fraction) as revealed by high speed centrifugation of spheroplasts lysate and they are absent in the chromosomal DNA fraction. dTTP 4-(N-2-chloroethyl-N-mehylamino) benzylamide suppresses completely the insensitive to actinomycin D action DNA synthesis and practically does not act on the sensitive one. It is suggested, that the DNA synthesis stable to the action of the antibiotic is catalyzed by RNA dependent DNA polymerase.