RNA polymerase isolated from bovine lymphosarcoma by sequential low- and high-salt extraction.
Bovine lymphosarcoma tissue has been extracted with low- and high-salt buffers [0.05 M Tris-C1 plus or minus 0.3 M (NH4) 2S04]. Diethylaminoethyl-Sephadex chromatography of both the high-salt and low-salt extracts yields RNA polymerases I and II, although low-salt extraction releases only one-third as much activity. Extraction by high salt of the residue from the low-salt extract, followed by diethylaminoethyl-Sephadex chromatography, yields additional enzyme activity with properties of Form II. Purification of the low-salt extract by protamine precipitation, elution with sodium succinate, and phosphocellulose chromatography yields a preparation of RNA polymerase (RNAP) with hybrid properties, combining the salt optimum of Form I, diethylaminoethyl-Sephadex elution pattern of form II, and alpha-amanitin sensitivity of Form III. RNAP. transcribes native D,A and chromatin efficiently. More RNAPL is recovered from lymphosarcoma tissue than from calf thymus.
D000546:Amanitins / Q000494:pharmacology; D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000201:enzymology; D011232:Chemical Precipitation; D002843:Chromatin; D002848:Chromatography, DEAE-Cellulose / Q000379:methods*; D012321:DNA-Directed RNA Polymerases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D066298:In Vitro Techniques; D008228:Lymphoma, Non-Hodgkin / Q000201:enzymology* / Q000662:veterinary; D011479:Protamines; D013386:Succinates; D013698:Templates, Genetic; D013950:Thymus Gland / Q000201:enzymology; D014158:Transcription, Genetic; D014316:Tritium
D000546:Amanitins; D002843:Chromatin; D011479:Protamines; D013386:Succinates; D014316:Tritium; D012321:DNA-Directed RNA Polymerases