Isolation of ribonucleic acid polymerases I, II, and III from Saccharomyces cerevisiae.
A procedure for the simultaneous purification of RNA polymerases I, II, and III from Saccharomyces cerevisiae is described. High yields of each enzyme activity are obtained, allowing the preparation of approximately 10 mg of polymerase I, 25 mg of polymerase II, and 12 mg of polymerase III from 1.2 kg of cells (wet weight). Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate indicates RNA polymerase I contains polypeptides with molecular weights 185 000, 137 000, 41 000, 35 000, 28 000, 24 000, 20 000, 16 000, 14 500, and 12 300; RNA polymerase II contains subunits with molecular weights 170 000, 145 000, 41 000, 33 500, 28 000, 24 000, 18 000, 14 500, and 12 500; and RNA polymerase III contains polypeptides with molecular weights 160 000, 128 000, 82 000, 53 000, 41 000, 37 000, 34 000, 28 000, 24 000, 20 000, 14 500, and 10 700.
D000546:Amanitins / Q000494:pharmacology; D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D046911:Macromolecular Substances; D008970:Molecular Weight; D009691:Nucleic Acid Denaturation; D012318:RNA Polymerase I / Q000302:isolation & purification*; D012319:RNA Polymerase II / Q000302:isolation & purification*; D012320:RNA Polymerase III / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
D000546:Amanitins; D046911:Macromolecular Substances; D012319:RNA Polymerase II; D012321:DNA-Directed RNA Polymerases; D012318:RNA Polymerase I; D012320:RNA Polymerase III