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. 2013 Jun 4;2:e00411. doi: 10.7554/eLife.00411

Figure 2. Termination complexes in association with RF1 alone and with both RF1 and apo-RF3.

(A) RC-RF1 (resolution 8.4 Å). (B) Close-up of RF1 in the map. (C) RF1 density from (B). Domains 1, 2/4 and 3 of RF1 are labeled. (D) RC–RF1•RF3 (resolution 9.7 Å). (E) Close-up of RF1and apo-RF3 in the map. RF1 contacts both L11—L11 contact point is labeled ‘L11 cp’— and apo-RF3. (F) Density of RF1 and RF3 from (E): RF1 domains labeled in black and RF3 domains 1, 2 and 3 in blue. (G)–(H) In the RC–RF1•RF3 complex, apo-RF3 does not interact with L11 or L6/SRL. Labels: 30S: small ribosomal subunit (yellow), 50S: large ribosomal subunit (blue), cp: central protuberance, L11: ribosomal protein L11, sp: spur, bk: beak, L1: L1 stalk, RF1: release factor 1, RF3: release factor 3, L6/SRL: position of ribosomal protein L6 and the sarcin/ricin loop, L11-NTD: ribosomal protein L11 N-terminal domain, arc: arc-like density. Density maps obtained from the total data set and of the RC-class not occupied by RF1 or RF3 are presented in Figure 2—figure supplement 1 and Figure 2—figure supplement 2, respectively. Additional panels of isolated RF1- and RF1, apo-RF3-densities are presented in Figure 2—figure supplement 3. FSC curves for resolution assessment of all density maps obtained can be found in Figure 2—figure supplement 4.

DOI: http://dx.doi.org/10.7554/eLife.00411.004

Figure 2.

Figure 2—figure supplement 1. Density map resulting from refinement of the full data set.

Figure 2—figure supplement 1.

Map is displayed at a resolution of 6.3 Å in accordance with a Fourier shell correlation (FSC) of 0.14 between half sets. Labels same as in Figure 2.
DOI: http://dx.doi.org/10.7554/eLife.00411.005
Figure 2—figure supplement 2. Density map resulting from refinement of the ‘RC’ class identified in our classification.

Figure 2—figure supplement 2.

The map shows the ribosome occupied with a P-site tRNA but no release factors.
DOI: http://dx.doi.org/10.7554/eLife.00411.006
Figure 2—figure supplement 3. Segmented RF1 and RF1, apo-RF3 densities.

Figure 2—figure supplement 3.

(A), (C) Segmented RF1 density from RC-RF1, compared to the segmented, combined RF1, apo-RF3 density from RC-RF1•RF3 (B), (D). Density marked by asterisk in (A) and (C) is no longer present in (B) and (D); instead redistributed density is observed in RF1 domain 1 (asterisk in [B] and [D]) upon apo-RF3 binding. The arc of RF3 is labeled in all panels featuring RF3. L11 cp marks the point of contact between L11 and RF3 in the RC-RF1•RF3 map. RF3 domains labeled in blue; RF1 domains in black.
DOI: http://dx.doi.org/10.7554/eLife.00411.007
Figure 2—figure supplement 4. FSC curves for density maps obtained from total data, RC, RC-RF1 and RC-RF1•RF3.

Figure 2—figure supplement 4.

Resolutions, determined using the FSC = 0.5 cutoff criterion, are 8.0 Å (full data set), 8.4 Å (RC-RF1), 9.7 Å (RC-RF1•RF3) and 11.8 Å (RC).
DOI: http://dx.doi.org/10.7554/eLife.00411.008