Open AccessStimulation of Eukaryotic DNA-Dependent RNA Polymerase by Protein Factors
Author(s) -
Sheng-Chung Lee,
Michael E. Dahmus
Publication year - 1973
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.70.5.1383
Subject(s) - rna polymerase , polymerase , rna , dna , microbiology and biotechnology , biology , escherichia coli , stimulation , biochemistry , chemistry , gene , neuroscience
Protein factors that stimulate DNA-dependent RNA polymerase activityin vitro have been purified from Novikoff ascites cells. These factors are not adsorbed by the diethylaminoethylcellulose column used for RNA polymerase purification and appear in the flow-through fraction. They can be fractionated into two classes by chromatography on carboxymethylcellulose. The first peak of activity elutes at 0.1 M NH4 Cl, is stable to heat treatment of 100° for 5 min, and is designated heat-stable factor; the second peak of activity elutes at 0.3 M NH4 Cl, is heat labile, and is designated heat-labile factor. Heat-labile factor can be further resolved into two components by chromatography on phosphocellulose. The heat-stable factor and second heat-labile factor stimulate the activity of (Novikoff ascites) RNA polymerase B by several-fold, and appear to function independently. RNA synthesis is stimulated only with native DNA as a template. No stimulation ofEscherichia coli RNA polymerase is observed with any of the factors.