title: Utilization of 5-alkyl UTPs by DNA-dependent RNA polymerase I and II purified from cherry salmon (Onchorhynchus masou) liver.
abstract: DNA dependent RNA polymerase II was purified to approximately 8300 fold from sonicated nuclear extract of cherry salmon (Onchorhynchus masou) liver by the following purification steps: polyethylene glycol treatment, DEAE=Sephadex A-25 column chromatography, heparin-Sepharose column chromatography, and affinity chromatography on DNA-cellulose. Final preparation of this enzyme has a specific activity of 157 nmole UMP incorporation into RNA per mg of protein per 10 min at 25 degrees. RNA polymerase I was also purified to approx. 3800 fold in a similar manner. Its specific activity was calculated as 26.2 nmole/mg/10 min. Utilization of various UTPs of these enzymes was studied by substitution experiments under the condition of limited synthesis. 5-Methyl UTP (rTTP) could be utilized by the RNA polymerase I 1.7 fold more efficiently compared with UTP. In contrast, the RNA polymerase II recognized rTTP as a substrate as efficiently as UTP. Similar experiments using other alkyl UTPs have been performed.
mesh_terms: D000478:Alkylation; D000818:Animals; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D008099:Liver / Q000201:enzymology*; D012318:RNA Polymerase I / Q000302:isolation & purification / Q000378:metabolism*; D012319:RNA Polymerase II / Q000302:isolation & purification / Q000378:metabolism*; D012474:Salmon; D013379:Substrate Specificity; D014500:Uracil Nucleotides; D014544:Uridine Triphosphate / Q000031:analogs & derivatives*
chemical_list: D014500:Uracil Nucleotides; D012319:RNA Polymerase II; D012321:DNA-Directed RNA Polymerases; D012318:RNA Polymerase I; D014544:Uridine Triphosphate