Figure 4.
Effects of cap analogs and SELEX RNAs on the formation of 80S ribosomal complexes on capped LUC mRNA molecules. (A) Formation of 80S ribosomal complexes in the presence of 1.2 mM cycloheximide (□), 1.2 mM cycloheximide and 5 mM m7-GTP (◊), or 1.2 mM cycloheximide and 5 mM EDTA (○) after sucrose gradient centrifugation is shown. The radioactivity in collected fractions is shown. The top of the gradient and the position of 80S ribosomes are indicated. (B) Formation of 80S ribosomal complexes in the absence of any inhibitor (○), in the presence of 8 mM GMP-PNP (▴), or 8 mM GMP-PNP and 5 mM m7-GTP (□) is depicted. (C) Formation of 80S complexes in the presence of 1.2 mM cycloheximide (◊), 1.2 mM cycloheximide and 1.5 mM (60-fold molar excess) of R8-35 (○), or 1.2 mM cycloheximide and 1.5 mM R0-1 (□) is shown. The conditions for translation in yeast extracts are the same as described in Fig. 3, except for the addition of translation inhibitors.