Characterization of vesicular stomatitis virus replicating complexes isolated in renografin gradients

Abstract

Newly synthesized products of replication and transcription from vesicular stomatitis virus (VSU)-infected HeLa cells have been separated on Renografin gradients and characterized. The replicative products have been identified by the following criteria: (1) sucrose-velocity gradients and methyl-mercury agarose gel analyses show RNA of sizes between 18 S and genome size 42 S. (2) This RNA is predominantly minus stranded by hybridization analyses. (3) A majority of the nascent RNA molecules of less than half genome size from replicative complexes are complementary to the 3′ end of the VSV plus strand, i.e., the L protein cistron. These short nascent RNA species are nuclease resistant and thus are presumed to be contained in ribonucleoprotein complexes.