Purification of mitochondrial RNA polymerase from Saccharomyces cerevisiae.
The RNA polymerase from the mitochondria of Saccharomyces cerevisiae has been extensively purified by Sepharose 4B, heparin Sepharose 4B phosphocellulose, and DEAE-Sephadex A-50 chromatography. The activity co-sediments with a 45,000-dalton polypeptide at 6.3 S in glycerol gradients. The activity is inhibited by antibodies to the 45,000-dalton polypeptide. The activity is not inhibited by rifampicin or alpha-amanitin. It requires Mg2+ and is inhibited by elevated ionic strength and Mn2+. The most efficient template for the RNA polymerase is poly[d(AT)], with mtDNA being the preferred natural template. The RNA polymerase transcribes mtDNA from the petite strain F11 in a nonrandom manner.
D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification* / Q000378:metabolism; D007118:Immunoassay; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D008928:Mitochondria / Q000201:enzymology*; D008970:Molecular Weight; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D014158:Transcription, Genetic
D008345:Manganese; D012321:DNA-Directed RNA Polymerases; D008274:Magnesium