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.