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. 2011 Mar 29;30(9):1804–1817. doi: 10.1038/emboj.2011.93

Figure 6.

Figure 6

Dissociation by Pelota, Hbs1 and ABCE1 of pre-termination complexes containing different numbers of mRNA nucleotides downstream of the P-site. (A) Structure of mRNAs encoding MVHC tetrapeptide followed by the UAG stop codon and containing 3, 6, 9, 13 or 17 nts after the last sense codon. (B, F) Ribosomal association of [35S-Cys]MVHC-peptidyl tRNA after incubation of pre-TCs assembled on mRNAs shown in panel (A) with (B) recombinant and (F) native Pelota, Hbs1 and ABCE1 at 1.5 mM Mg2+, assayed by native gel electrophoresis. (C) Ribosomal association of [32P]MVHC-9 mRNA after incubation of ECs with Pelota, Hbs1 and ABCE1 at 1.5 mM free Mg2+, assayed by native gel electrophoresis. (D) Purified native and recombinant Pelota resolved by SDS–PAGE. (E) Dissociation by native Pelota, Hbs1 and ABCE1 at 1.5 mM free Mg2+ of pre-TCs assembled on MVHL-STOP mRNA using [32P]60S subunits depending on their treatment with RelE, assayed by SDG centrifugation. Upper fractions were omitted for clarity.