Figure 8
Extra 3CDpro(3CproH40A) has no effect on virus production and RNA synthesis in reactions programmed with PV transcript RNA. Translation RNA-replication reactions were carried out, as described in Materials and Methods. The viral RNA template was replaced with a PV full-length transcript RNA made from a T7 promoter or with a ribozyme-treated transcript RNA. Where indicated 3CDpro(3CproH40A) (5.5 nM) was added at t = 0 hr. (A) Comparison of virus yields in reactions templated with viral RNA and transcript RNAs. (B) Plus strand RNA synthesis with initiation complexes isolated from reactions programmed with PV transcript RNA made from a T7 promoter (Materials and Methods). Lane 1, CRC: [32P]-labeled RNA products obtained from crude replication complexes (Materials and Methods). (C) Plus strand RNA synthesis with initiation complexes isolated from reactions programmed with ribozyme-treated PV transcript RNA (R+ RNA). Lane 1, CRC: [32P]-labeled RNA products obtained from crude replication complexes (Materials and Methods).