title: Daunorubicin inhibition of DNA-dependent RNA polymerases from Ehrlich ascites tumor cells.
abstract: Two different forms of DNA-dependent RNA polymerase have been solubilized and purified from nuclei of Ehrlich ascites tumor cells. The purification procedure involves ammonium sulfate precipitation and gel filtration on Sephadex G-25. The separation of A and B activities is achieved by chromatography on DEAE-cellulose. Nuclei are prepared from cells, sensitive or resistant to daunorubicin. RNA polymerases A and B have an absolute requirement of divalent cations for activity. Native DNAs are better templates than heat-denatured DNAs for RNA polymerase A. On the contrary heat-denatured DNA is more transcribed than the native one by RNA polymerase B. The low level of transcription of total and nucleolar ascites DNAs is due to the DNA, the same results being obtained with ascites and calf thymus RNA polymerases A and B. The inhibitory action of daunorubicin on RNA polymerases A and B from Ehrlich ascites tumor cells has been studied in vitro. The same results are obtained with enzymes extracted from sensitive or resistant cells. Daunorubicin does not inhibit the binding of RNA polymerases to the DNA template, but prevents the transformation of the DNA-daunorubicin-RNA-polymerase unstable complex into the highly stable one. This inactive ternary complex has a dissociation rate faster than the stable complex formed without daunorubicin. The size of the RNA synthesized in the presence or absence of daunorubicin is the same.
mesh_terms: D000818:Animals; D002286:Carcinoma, Ehrlich Tumor / Q000201:enzymology*; D002466:Cell Nucleolus / Q000187:drug effects / Q000201:enzymology; D002467:Cell Nucleus / Q000187:drug effects / Q000201:enzymology; D012321:DNA-Directed RNA Polymerases / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D003630:Daunorubicin / Q000494:pharmacology*; D004351:Drug Resistance; D005260:Female; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D008433:Mathematics; D051379:Mice; D009994:Osmolar Concentration; D013698:Templates, Genetic; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D008345:Manganese; D012321:DNA-Directed RNA Polymerases; D008274:Magnesium; D003630:Daunorubicin