In vitro synthesis of genome length complementary RNA of vesicular stomatitis virus in the presence of inosine 5′-triphosphate

Abstract

The virion-associated RNA polymerase of vesicular stomatitis virus (VSV) synthesizes RNA in vitro when GTP is replaced by inosine 5′-triphosphate (ITP). The synthesis is optimal at an ITP concentration of 200 μ M and the extent of synthesis is between 15 to 20% compared to normal transcription in the presence of GTP. Analyses of the RNA products revealed that approximately 10% of the product RNA represented plus-strand complement of the genome RNA. Defective interfering particles of VSV were also capable of synthesizing complementary RNA in the presence of ITP, in addition to 46-base RNA, although in lesser amount (2%). Since I substitution facilitates read-through of the genome RNA, these results suggest that interaction of the product RNA with the genome template or ITP-mediated modification of a viral protein may be involved in the readthrough process in vitro.