Kinetics of appearance of the products of poliovirus-induced RNA polymerase

Abstract

Polyacrylamide gel electrophoresis has been utilized to analyze the products of the membrane-associated RNA polymerase induced in HeLa cells by poliovirus infection. Single-stranded viral RNA as well as double-stranded RNA (RF) and the multistranded replicative intermediate (RI) have been identified by virtue of their characteristic electrophoretic mobility and other criteria. Kinetic studies including a “pulse-chase” experiment have provided unequivocal evidence that RI represents the functional intermediate in the synthesis of single-stranded viral RNA. Single-stranded RNA does not appear as a product of enzyme preparations rendered soluble by sodium deoxycholate and addition of deoxycholate to a membrane-associated preparation during the course of the reaction leads to the degradation of previously synthesized single-stranded RNA. Study of the degradation of added single-stranded RNA suggests that the phenomenon is probably the consequence of the accessibility of the products of the soluble enzyme to nucleases in the preparation.