Journal:

Article Title: Ribosome hijacking: a role for small protein B during trans -translation

doi: 10.1038/embor.2008.243

Figure Lengend Snippet: Mapping of accessible Gs and As of Escherichia coli 16S ribosomal RNA within a stalled ribosome. (A) Kethoxal and (B) DMS. (Top) Autoradiograms of 12% polyacrylamide gels of the cleavage products of extended cDNAs. Lanes U, G, C and A: sequencing ladders. (Bottom) Quantification of the protections. (C) Immunoblotting of stalling ‘truncated mRNA–tRNAfMet–30S–SmpB' complexes from wt 30S or mutants at positions 530, 1492 and 1493. (D) Equilibrium dissociation experiments of tmRNA on stalled ribosomes from filter-binding assays, with maximum binding set to 1 and the other values expressed as the bound fraction: stalled 70S pre-loaded with wt SmpB (diamonds) or with Δ-Cter SmpB (squares). DMS, dimethylsulphate; mRNA, messenger RNA; SmpB, small protein B; tRNA, transfer RNA; tmRNA, transfer messenger RNA; wt, wild type.

Article Snippet: The 30S mutants were provided by Dr R. Green ( Johns Hopkins University, USA).

Techniques: Sequencing, Western Blot, Binding Assay

Journal:

Article Title: Ribosome hijacking: a role for small protein B during trans -translation

doi: 10.1038/embor.2008.243

Figure Lengend Snippet: Model of the decoding centre–SmpB interaction within the ribosome. 50S and 30S subunit surfaces from a Thermus thermophilus ribosome are shown in cyan and pale green, respectively; E-site and P-site tRNAs in red and orange; mRNA in blue; TLD in grey; and SmpB in blue. The parts of h44 and h18 involved in the decoding are shown in ribbon mode (pink and yellow). Adenines 1408, 1492, 1493 from h44 and G530 from h18 are in red. BR2 SmpB amino acids are coloured as in Fig 2. (Inset) Comparison with the IF1 structure (purple) with the decoding site (pink, PDB 1HR0). Red: decoding centre A1408, A1492 and A1493; cyan: R41 and R46 stack against the A1492 and A1493, respectively. BR2, binding region 2; IF1, Initiation factor 1; SmpB, small protein B.

Article Snippet: The 30S mutants were provided by Dr R. Green ( Johns Hopkins University, USA).

Techniques: Comparison, Binding Assay