Comparison of replication complexes synthesizing poliovirus RNA

Abstract

Poliovirus RNA replication complexes prepared by homogenization or detergent-extraction of infected cells have been used to study RNA replication in vitro. We show that the products are different depending on the method of extraction. Complexes derived from homogenized cells synthesize full-length single-stranded poliovirus RNA, replicative intermediate (RI), and replicative form (RF) RNA species, whereas only RI and RF are made by detergent-extracted complexes. Both complexes synthesize predominantly plus-strand strand RNA in vitro. Detergent treatment of complexes from homogenized cells inhibits synthesis of full-length, single-stranded RNA. The data suggest that the poliovirus replication complex contains several enzymatic activities and that the ability to synthesize full-length single-stranded RNA requires the association of a labile, possibly membrane associated, factor with the core elongation complex. Furthermore, RNase T 1-resistant oligonucleotide fingerprints of the products of detergent-extracted complexes are missing a specific internal oligonucleotide, which has been mapped near, but not identical to, oligonucleotides missing in fingerprints of DI virus genome.