Studies of the Ribonucleic Acid Polymerase from Escherichia coli

Detailed studies (l-6) of ribonucleic acid polymerase have demonstrated the role of deoxyribonucleic acid in serving as the template for ribonucleic acid synt,hesis.Partially purified preparations of RNA polymerase from Sficrococcus Zysodeikticus have also been shown (7, 81 to catalyze RNA synthesis in the presence of natural and synthetic Rn'l templates.Similar studies have been reported(9) with partially purified RNA polymerase preparations from Azotobacter vinelandii.Both groups of investigators have concluded that the RNA-directed reaction is catalyzed by RNA polymerase.Previous publications (3, 4) from this laboratory noted that with partially purified RNA polymerase from Escherichia COG, synthetic polyribonucleotides serve efficiently as templates for the synthesis of complementary polyribonucleotides.This paper reports in greater detail some of the properties of polyribonucleotide-directed reactions.Evidence will be presented to show that the same enzyme, RNA polymerase, is involved in the synthesis of RNA with the use of either DF-\ or synthetic polyribonucleotides as templates.EXPERIMENTAL PROCEDURE Materials and Methods--ATP-8-W, GTP-8-'4C, CTP-2-W, and UTP-2-14C were obtained from Schwarz BioResearch, Inc., and were purified by paper chromatography in several solvents before use.ATP, CTP, GTP, and GTP were obtained from Pabst Laboratories and, if necessary, purified by paper chromatography before use.Poly A,] poly U, poly C, and poly CU (base ratios, 1O:l and 1: 10) were obtained from Miles Laboratories.Poly A, poly U, and poly C were found to be homogeneous during ultracentrifugation in 0.15 M NaCl-0.015M sodium citrate buffer, pH 7.1, and the s20,W values were calculated to be about 9.9, 5.4, and 4.0, respectively.Poly AU (base ratio, 1: 1) was kindly furnished by Dr. Alexander Rich.Bacteriophage Tz DNA was prepared according to the procedure of Mandell and Hershey (10).Tz DNA was heated according to the procedure of Marmur and