The 24,000 Da subunit is not required for the RNA synthesis activity of chicken leukemia RNA polymerase II.
The partially purified RNA polymerase II from chicken leukemia cells (Chuang R. Y., Chuang L. F. & Israel M. (1986) Biochem. Pharmacol. 35, 1293-1297) contained multiple subunits with molecular masses (in Da) ranging from 220,000 to 24,000, as shown by SDS-polyacrylamide gel electrophoresis. The enzyme was further purified through phosphocellulose column and fractions containing the enzyme activity were collected and concentrated 400-fold through a microconcentrator. The microconcentrator contained a membrane with a molecular weight cutoff around 30,000 and, hence, removed the 24,000 Da polypeptide from the enzyme. It was found that the resulting enzyme retained all the catalytic activity as compared to the enzyme preparation before the concentration step, suggesting that the stoichiometric amount of the 24,000 Da polypeptide is not required for RNA synthesis activity with a denatured DNA template.
Chuang
L F
LF
Department of Pharmacology, School of Medicine, University of California, Davis 95616.
Chuang
R Y
RY
D000818:Animals; D001353:Avian Leukosis / Q000201:enzymology; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004795:Enzyme Stability; D012319:RNA Polymerase II / Q000378:metabolism*; D014462:Ultrafiltration
D012319:RNA Polymerase II