Full-length viral rna synthesized in vitro by vesicular stomatitis virus-infected hela cell extracts

Abstract

Total cell extracts from vesicular stomatitis virus (VSV)-infected HeLa cells and purified vesicular stomatitis virions were assayed for their abilities to synthesize RNA in vitro. Analysis of the products of these reactions by methylmercury-agarose gel electrophoresis showed that the infected cell extracts directed the synthesis of 42 S RNA as well as of intact VSV-specific messenger RNAs. The majority of the 42 S RNA synthesized by the infected cell extract bound to an oligo(dT)-cellulose column. Extracts prepared from infected cells treated with cycloheximide produced no detectable 42 S RNA, suggesting that this product results from viral RNA replication rather than transcription. When viral ribonucleoprotein cores were purified from the infected cell extract and assayed in vitro, synthesis of 42 S RNA was markedly decreased. As previously reported, purified virions produced no 42 S RNA but directed the synthesis only of intact messenger RNAs and a small RNA which resembles the previously described “leader” RNA. Additional species of RNA were synthesized in the in vitro systems containing virions or cell extracts; these VSV-specific RNAs have not been observed in previous studies and their function and composition is unknown.