Q-beta-replicase and E. coli transcriptase: requirements for substrate selection as revealed by a study of base...

Base selection by the Qbeta-replicase and E. coli transcriptase has been studied using substrate analogs. Six analogs with normal hydrogen-bonding sites were polymerized by both enzymes. Three analogs containing ring substitutions which affect the conformation at the glycosyl bond would not substitute for their normal congeners. The remaining analog was an excellent substrate for the transcriptase but not the replicase.The results imply that the base selection procedure has stringent requirements for substrate conformation. Part of the restriction may derive from the requirement that template and substrates conform to the stereochemical constraints of a double helix. In the Qbeta-replicase reaction, synthesis in the presence of substrate analogs displayed abortive kinetics, implying that the replicase reaction can be uniquely curtailed by substrate analogs.