Studies on the product binding sites of the Azotobacter vinelandii ribonucleic acid polymerase.
During chain elongation RNA polymerase exists as a ternary DNA-enzyme-RNA complex in which a discrete length of the nascent RNA chain proximal to the 3'-OH terminus will be bound to the product binding site (Krakow, J. S., and Fronk, E. (1969) J. Biol. Chem. 244, 5988). We have utilized the poly[d(A-T)]-directed reaction to determine the length of the nascent poly[r(A-U)] protected from attack by pancreatic ribonuclease. Following release of the ribonuclease resistant oligo[r(A-U)] from the ternary complex, its size was determined by ion exchange chromatography on DEAE-cellulose, gel filtration on Bio-Gel P-10, and the ratio of 3'-terminal uridine to internal 2':3'-UMP following alkaline hydrolysis. The results indicate that the length of the nascent protected fragment is approximately 12 residues.
D001395:Azotobacter / Q000201:enzymology*; D001665:Binding Sites; D002848:Chromatography, DEAE-Cellulose; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D007700:Kinetics; D009841:Oligonucleotides / Q000032:analysis; D011119:Polynucleotides; D011485:Protein Binding; D013698:Templates, Genetic; D013997:Time Factors; D014158:Transcription, Genetic
D009841:Oligonucleotides; D011119:Polynucleotides; D012321:DNA-Directed RNA Polymerases