RNA polymerases from avian liver. Isolation and fractionation by heparin-sepharose chromatography.
Optimal conditions for isolation of rooster liver nuclei were studied for the purification of RNA polymerases I and II by varying the concentrations of sucrose, magnesium chloride and spermine in the isolation medium. Addition of spermine and/or magnesium chloride to the hypertonic sucrose medium was found to be advantageous for the purification. Heparin-Sepharose chromatography can be recommended for purification of RNA polymerases when used in a stepwise manner. Furthermore, gradient elution in the heparin-Sepharose chromatography was found to separate RNA polymerases I and II. RNA polymerase III was eluted with RNA polymerase I. A few minor peaks of RNA polymerase II activity were detected with gradient elution. Factors influencing the affinity of RNA polymerases towards heparin-Sepharose are discussed.
D000818:Animals; D002467:Cell Nucleus / Q000201:enzymology; D002847:Chromatography, Agarose / Q000379:methods*; D002850:Chromatography, Gel / Q000379:methods*; D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification*; D006493:Heparin; D008099:Liver / Q000166:cytology / Q000201:enzymology*; D011200:Poultry / Q000378:metabolism*; D012318:RNA Polymerase I / Q000302:isolation & purification; D012319:RNA Polymerase II / Q000302:isolation & purification
D006493:Heparin; D012319:RNA Polymerase II; D012321:DNA-Directed RNA Polymerases; D012318:RNA Polymerase I