The Path of Messenger RNA through the Ribosome

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The Path of Messenger RNA through the Ribosome Gulnara Zh. Yusupova, Marat M. Yusupov, J.H.D. Cate, Harry F. Noller  Cell  Volume 106, Issue 2, Pages 233-241 (July 2001) DOI: 10.1016/S0092-8674(01)00435-4

Figure 1 Nucleotide Sequences of the Three Model mRNAs Used in this Study The Shine-Dalgarno sequence (S/D), and P- and A-site codons are underlined. The self-complementary sequences forming the putative A-site helix in MF36 mRNA are overlined Cell 2001 106, 233-241DOI: (10.1016/S0092-8674(01)00435-4)

Figure 2 Fourier Difference Maps of mRNAs (A) 7 Å Fourier difference map of MK27 mRNA with the mRNA model (yellow) docked, showing the position of the Shine-Dalgarno (S/D) helix (magenta) and the positions of the A- and P-site codons (orange and red, respectively), viewed from the top of the 30S ribosomal subunit. (B) Difference map of the MF36 mRNA, showing a four-base-pair tetraloop helix (A-site helix) fitted to the extra density at the A site. (C) Same as for (B), except that the A-tRNA anticodon stem-loop (green) is shown in the position observed experimentally in the A-tRNA difference map (Yusupov et al., 2001), in place of the A-site mRNA helix. The five-nucleotide (GGAGG/CCUCC) core of the Shine-Dalgarno interaction is shown in magenta, and the rest of the 16S rRNA tail in cyan Cell 2001 106, 233-241DOI: (10.1016/S0092-8674(01)00435-4)

Figure 3 Path of mRNA through the 30S Ribosomal Subunit (A) Interface and (B) solvent views of the mRNA in the 30S ribosomal subunit. A, P; the A- and P-site codons. 5′, 3′; the 5′ and 3′ correspond to positions −15 and +15 of the mRNA model. The head, platform, shoulder, and body of the subunit, and ribosomal proteins S2, S3, S4, S5, S7, S11, S12, and S18 are indicated. The ribosomal proteins are shown in dark blue, 16S rRNA in cyan and the mRNA is colored as in Figure 2 Cell 2001 106, 233-241DOI: (10.1016/S0092-8674(01)00435-4)

Figure 4 Detailed Views of mRNA-Ribosome Interactions (A) Solvent-side stereo view of the Shine-Dalgarno (S/D) helix bound in its cleft, formed by helices 20, 28, and 37 (h20, h28, h37) and the 723 loop of 16S rRNA and proteins S11 and S18, and the path of mRNA nucleotides −1 to −4 through the upstream tunnel. The MV36 Fourier difference map is shown. (B) Interface stereo view of the A- and P-site codons and their flanking nucleotides (530, 790, 791, 926, 1492, 1493, and 1498) in 16S rRNA. The locations of restrictive mutations in ribosomal protein S12 are shown in yellow (the universal PNSA sequence) and orange. The positions of the bases for G926 and U1498 are modeled from the high-resolution structure of the T. thermophilus 30S subunit (Wimberly et al., 2000). (C) Same as (B), but with the A- and P-tRNAs (orange and red, respectively) docked according to their experimentally observed locations (Yusupov et al., 2001) Cell 2001 106, 233-241DOI: (10.1016/S0092-8674(01)00435-4)

Figure 5 The Downstream Tunnel (A) Interface stereo view of the downstream tunnel, showing the features of 16S rRNA layer surrounding mRNA positions +7 to +10. (B) Solvent-side stereo view of the downstream tunnel, showing the formation of the protein layer surrounding positions +11 to +15 of the mRNA by proteins S3, S4, and S5. The double-stranded RNA binding domain of protein S5 is shown in magenta. The MK27 difference map is shown Cell 2001 106, 233-241DOI: (10.1016/S0092-8674(01)00435-4)

Figure 6 Arrangement of Ribosomes and mRNA in the Crystal View down the crystallographic 4-fold axis of the 70S ribosome-mRNA-tRNA complex (Yusupov et al., 2001), showing the head-to-tail juxtaposition of the model mRNAs (red-orange) between adjacent ribosomes. The molecular components shown are 16S rRNA (cyan), 23S rRNA (gray), 5S rRNA (gray-blue), small subunit proteins (dark blue), large subunit proteins (magenta), the A-, P-, and E-tRNAs (yellow, orange, and red), and the mRNA (red-orange) Cell 2001 106, 233-241DOI: (10.1016/S0092-8674(01)00435-4)