Purification and characterization of RNA polymerase I from a higher plant.
RNA polymerase I was purified from chromatin isolated from auxin-treated soybean hypocotyl. Purification was achieved by using Agarose A-1.5m gel filtration, DEAE-cellulose, CM-sephadex, and phosphocellulose chromatography, and sucrose density  gradient centrifugation. With denatured calf thymus DNA as template, the enzyme has a high specific activity (200-300 nmol/mg/30 min at 28 degrees C) which is comparable to other RNA polymerase I enzymes purified from animals and yeast. While the gel profiles indicate that purification to homogeneity (greater than 90%) may not have been achieved, the enzyme appears to be composed of possibly 7 subunits, several of which are similar to the subunits of yeast RNA polymerase I. The putative subunits and molar ratios are 183 000 (1), 136 000 (1), 50 000 (0.5), 46 000 (0.5), 40 000 (0.5), 33 000 (0.2), and 28 000 (2). The purified enzyme strongly prefers a completely denatured template such as poly(dC).
D000546:Amanitins / Q000494:pharmacology; D000645:Ammonium Sulfate / Q000494:pharmacology; D002843:Chromatin / Q000302:isolation & purification; D003513:Cycloheximide / Q000494:pharmacology; D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification* / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D010944:Plants / Q000201:enzymology*; D011119:Polynucleotides / Q000494:pharmacology; D012293:Rifampin / Q000494:pharmacology; D013698:Templates, Genetic
D000546:Amanitins; D002843:Chromatin; D011119:Polynucleotides; D008345:Manganese; D003513:Cycloheximide; D012321:DNA-Directed RNA Polymerases; D008274:Magnesium; D000645:Ammonium Sulfate; D012293:Rifampin