Fig 1. Release of tRNA during the preparation of the PoTC as schematically described in S1 Fig.

Various amounts of polysomes were incubated with 2 μM EF-G and 1 mM GTP and isolated (A in S1 Fig). Total reaction mixture was 50 μl. The polysomes (B in S1 Fig) thus isolated were incubated with 50 μM puromycin in buffer R. The tRNA released during these operations was isolated by passing through nitrocellulose membrane (0.45 nm). Transfer RNA thus obtained was measured by aminoacylation with a mixture of [¹⁴C] amino acids. The Y-axis shows the released tRNA (pmols) calculated from [¹⁴C]-aminoacyl-tRNA (cpm) as described in Materials and methods and the supplement. Squares, tRNA released by EF-G/GTP; circles, tRNA released by puromycin and EF-G/GTP.