Fig. (4) (a) Translational activation of encapsidated AltMV-MU RNA in vitro. SDS-PAGE in 8-20% polyacrylamide gel of ³⁵S-labeled translation products in the wheat germ extract. Gel was dried and autoradiographed. Translation of purified AltMV-MU RNA (lane 1); AltMV-MU virions (lane 2); AltMV-MU virions after phosphorylation with PKC (lane 3), AltMV-MU preparations incubated with AltMV-MU TGB1 (lane 4). AltMV-MU RNA concentration in the translation sample was 40 µg/ml. Arrows indicate the positions of molecular weight markers and the bold arrow indicates the position of the protein encoded by the 5'- proximal AltMV-MU replicase gene (174 kDa). (b) Far-western blot (Far-WB) analysis of interaction of different TGB1 proteins with CP AltMV-MU at pH 8.0. Purified TGB1 PAMV (1 µg, lane 1, negative control) and TGB1 AltMV-MU (1 µg, lane 2) proteins were separated by 8-20% SDS-PAGE. TGB1s were transferred to PVDF membrane, incubated with CP AltMV-MU (5µg/ml), mouse AS to CP AltMV-MU (dilution 1:10000) and anti-mouse antibodies conjugated with horseradish peroxidase (Promega); reaction was visualized by the ECL system (Amersham Biosciences).