Initiation of RNA synthesis in vitro by vesicular stomatitis virus: single internal initiation in the presence of aurintricarboxylic acid and vanadyl ribonucleoside complexes.

Abstract

In the presence of aurintricarboxylic acid (ATA) and vanadyl ribonucleoside complexes (VRC), we have isolated and characterized a small RNA, product of VSV in vitro transcription. This RNA is capped and lacks poly(A) at its 3'-end. Nucleotide sequence analysis revealed that this RNA corresponds to the 5'-terminal transcription product of the N-gene. The termination of the transcription occurs precisely at the 118th base from the 3'-end of the VSV genome. Analysis of the nucleotide sequence around this region reveals a potential secondary structure. Photoreaction of the VSV with 4'-substituted psoralen fails to inhibit the synthesis of the 68-mer RNA under conditions where full length mRNA synthesis is blocked, indicating that the psoralen binding site is located further into the N-gene. Since this RNA is the only in vitro transcription product synthesized under these conditions, the existence of two types of polymerase activities, one for the synthesis of leader RNA and one for mRNA, is suggested.