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. 2006 Mar 1;20(5):624–636. doi: 10.1101/gad.1397906

Figure 6.

Figure 6.

Influence of eIF5-induced hydrolysis of eIF2-bound GTP on 48S complex formation on AUG triplets in good and bad context (A,B) and on UV cross-linking of eIF2α to the −3 nucleotide of mRNA in 48S complexes (C,D). (A,B) Toe-print analysis of 48S complexes assembled on (CAA)n-AUGbad/bad-GUS mRNA from 40S subunits, Met-tRNAMeti, and eIFs as indicated. Toe-prints due to 48S complexes are shown on the left. (C,D) UV cross-linking of 32P-labeled (CAA)n-AUG-(CAA)m mRNA derivatives containing 4-thioU or 6-thioG at [−3] with components of 48S complexes before and after incubation with eIF5, eIF5B, and 60S subunits, as indicated. In lanes 2 48S complexes incubated with eIF5 were subjected to sucrose density gradient centrifugation before UV cross-linking. Cross-linked proteins were assayed by SDS-PAGE and autoradiography. eIF2α and rpS5 are indicated on the right.