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

Figure 5.

Figure 5.

Activities of α-subunit- and β-subunit-deficient eIF2 in 48S complex formation. (A) Sequence of the 5′-UTR of (CAA)n-AUGbad/bad-GUS, (CAA)n-AUGgood/bad-GUS, and (CAA)n-AUGbad/good-GUS mRNAs with initiation codons in bold. Context residues −3 to +4 are underlined. (B) Coomassie-stained SDS-PAGE with resolved complete, α-subunit- and β-subunit-deficient eIF2, and recombinant eIF2α. The positions of eIF2 subunits are indicated on the right. (C,F,G) Toe-print analysis of 48S complexes assembled on mRNAs (shown in A) from 40S subunits, Met-tRNAMeti, complete, α-subunit- and β-subunit-deficient eIF2, and other eIFs as indicated. Full-length cDNAs and toe-prints due to 48S complexes are shown on the side. (D,E) UV cross-linking of 32P-labeled (CAA)n-AUG-(CAA)m mRNA derivatives containing 4-thioU or 6-thioG at [−3] in 48S complexes assembled using forms of eIF2 as indicated, assayed by SDS-PAGE and autoradiography. eIF2α and rpS5 are indicated on the right.