Initiation of RNA synthesis in vitro by vesicular stomatitis virus. Role of ATP

Abstract

The ATP requirement during RNA transcription by the RNA-dependent RNA polymerase associated with purified vesicular stomatitis virus was studied during chain initiation and chain elongation steps. Initiation of transcription in vitro has an apparent Km for ATP of approximately 500 micron, whereas the apparent Km for ATP during chain elongation is almost identical with those for the other three ribonucleoside triphosphates, i.e. 20 to 30 micron. Further analysis of the transcription process demonstrated that the beta-gamma imido analogue of ATP cannot be used for the initiation step. However, it can be incorporated into RNA during chain elongation. These results indicate that a hydrolyzable form of the beta-gamma phosphodiester bond in ATP is required for the initiation process and also support the hypothesis for a single promoter site for the initiation of transcription on the genome of vesicular stomatitis virus.