Figure 3.

eIF2A facilitates binding of Met-tRNA_i to 40S ribosome in a HCV IRES-dependent manner. The binding of [³⁵S]Met-tRNA_i to 40S ribosome was monitored by a filter-binding assay. The requirement of HCV IRES RNA and eIF2A was tested by leave-one-out approach. Recombinant His–eIF2A protein purified from E. coli, 40S ribosomal subunit purified from HeLa S3 cell, HCV IRES RNA synthesized by T7 RNA polymerase, and [³⁵S]Met-tRNA_i charged by methionyl tRNA synthetase (Pestova and Hellen, 2005) were used in the filter-binding assays. Each experiment was performed three times. Columns and bars represent mean values±s.d. values.