FIG. 8.

Comparison of [³⁵S]methionine- and formyl-[³⁵S]methionine-labeled products of translation of HCV RNA in Krebs-2 S10. (A) Control translation of EMCV RNA showing that the f[³⁵S]Met-preparation used specifically labels only the N-terminal protein moieties in the viral polyprotein. The reaction mixtures containing either [³⁵S]methionine (lane 2) or f[³⁵S]Met-(lane 4) were incubated with EMCV RNA (20 μg/ml) at 34°C for 90 min, as described previously (74). Analyses of the samples that did not receive the exogenous mRNA and were incubated with either [³⁵S]methionine or f[³⁵S]Met-are shown in lanes 1 and 3, respectively. (B) HCV RNA (20 μg/ml) was translated for the times indicated in the absence (−; lanes 6 to 8) or presence (+; lanes 10 to 12) of CMMs, using f[³⁵S]Met-as a substrate. [³⁵S]methionine-labeled HCV RNA translation products synthesized after the indicated time intervals are shown in lanes 2 to 4 for comparison. In lanes 1, 5, and 9, no HCV RNA was added and the reaction mixtures were incubated for 180 min. The samples were analyzed by SDS-15% PAGE and autoradiography. The positions of virus-specific proteins are indicated on the right. Arrows indicate formyl-[³⁵S]methionine-labeled products.