Fig. 3.
Association of Dom34 and Hbs1 with the ribosome. (A) Association of Hbs1 with ribosomes in the presence of Dom34 and various nucleotides was visualized by Western blot analysis following pelleting of ribosomes through a sucrose cushion. Unlabeled Hbs1 was used as a chase to distinguish between transient and stable interactions. (B) Similar to A, looking at the interaction of Dom34 with ribosomes under various conditions by using an unlabeled Dom34 chase. (C) Association of Dom34 with ribosomes in the presence or absence of Rli1 and various nucleotides. Visualized as above and using an unlabeled Dom34 chase. (D) kcat and K1/2 determinations for Rli1-mediated subunit dissociation. Subunit dissociation reactions were performed at various concentrations of Rli1 in the presence or absence of Dom34. Subunit dissociation was monitored by formation of peptidyl-tRNA via native gel electrophoresis (n = 3, ± SEM).