Cryo-EM models

The cryo-EM revolution is sweeping structural biology. With the advent of the direct electron detector, cryo-EM (cryogenic electron microscopy) resolution rivals that of X-ray crystallography. We are automating the process of producing atomistic models from 3-D cryo-EM reconstructions.

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Ribosome Model 1

Ratje, A.H, Loerke, J, Mikolajka, A, Brünner, M, Hildebrand, P.W, Starosta, A.L, Dönhöfer, A, Connell, S.R, Fucini, P, Mielke, T, Whitford, P.C, Onuchic, J.N, Yu, Y, Sanbonmatsu, K.Y, Hartmann, R.K, Penczek, P.A, Wilson, D.N, and Spahn, C.M. (2010) Head Swivel on the Ribosome Facilitates Translocation by Means of Intra-Subunit tRNA Hybrid Sites. Nature 468 (7324), 713-716, published December 2, 2010.

All-atom model for cryoelectron microscopy maps of TIPRE (a, b) of the 70S–EF-G–GDP–FA complex are shown in mesh form with docked models in ribbon representation: EF-G (red), tRNA (green), 23S/5S rRNA (blue), 50S ribosomal proteins (orange), 16S rRNA (yellow) and the 30S ribosomal proteins (magenta). The maps are shown from the 30S side with the 30S subunit computationally removed (a) and from the 50S side with the 50S subunit computationally removed (b). CP, central protuberance; bk, beak; sp, spur; pt, platform; h, head; b, back; L1 and L7, ribosomal proteins L1 and L7, respectively; H69, helix 69 of 23S rRNA.

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Ribosome Model 2

Ratje, A.H, Loerke, J, Mikolajka, A, Brünner, M, Hildebrand, P.W, Starosta, A.L, Dönhöfer, A, Connell, S.R, Fucini, P, Mielke, T, Whitford, P.C, Onuchic, J.N, Yu, Y, Sanbonmatsu, K.Y, Hartmann, R.K, Penczek, P.A, Wilson, D.N, and Spahn, C.M.T. (2010) Head Swivel on the Ribosome Facilitates Translocation by Means of Intra-Subunit tRNA Hybrid Sites. Nature 468 (7324), 713-716, published December 2, 2010.

All-atom model for cryoelectron microscopy maps of TIPOST (c, d) of the 70S–EF-G–GDP–FA complex are shown in mesh form with docked models in ribbon representation: EF-G (red), tRNA (green), 23S/5S rRNA (blue), 50S ribosomal proteins (orange), 16S rRNA (yellow) and the 30S ribosomal proteins (magenta). The maps are shown from the 30S side with the 30S subunit computationally removed (c) and from the 50S side with the 50S subunit computationally removed (d). CP, central protuberance; bk, beak; sp, spur; pt, platform; h, head; b, back; L1 and L7, ribosomal proteins L1 and L7, respectively; H69, helix 69 of 23S rRNA.

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Ribosome Model 3

Whitford, P.C, Ahmed, A, Yu, Y, Hennelly, S.P, Tama, F, Spahn, C.M.T, Onuchic, J.N, and Sanbonmatsu, K.Y. (2011) Excited States of Ribosome Translocation Revealed Through Integrative Molecular Modeling. Proceedings of the National Academy of Sciences 108 (47), 18943-18948, published November 11, 2011. doi:10.1073/pnas.1108363108

Suggested models for transient configurations during translocation. Atomic model of the ribosome with tRNA molecules in a A/P*-P/E configuration, fitted to a cryo-EM density where the mRNA has not translocated (TIpre configuration). The A-site tRNA and domain IV of EF-G are not fit to the density. In the cryo-EM construct domain IV can enter the A site because there is no A/A tRNA present. Here, domain IV is not fit, but it is allowed to relax into an accessible low-energy configuration.

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Ribosome Model 4

Whitford, P.C, Ahmed, A, Yu, Y, Hennelly, S.P, Tama, F, Spahn, C.M.T, Onuchic, J.N, and Sanbonmatsu, K.Y. (2011) Excited States of Ribosome Translocation Revealed Through Integrative Molecular Modeling. Proceedings of the National Academy of Sciences 108 (47), 18943-18948, published November 11, 2011. doi:10.1073/pnas.1108363108

Suggested models for transient configurations during translocation. Atomic model of the ribosome and tRNAs in an A/P-P/E configuration, fitted to a density map in which the mRNA has partially translocated (TIpost). The concomitant displacement of the A-site tRNA and partial mRNA translocation leads to available space for fitting domain IV of EF-G. Therefore, all atoms were fit to the density, except for the A/P tRNA.

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Sanbonmatsu Team
Los Alamos National Laboratory
Theoretical Biology and Biophysics Group
P.O. Box 1663, Los Alamos, NM 87545
Public Affairs Office: (505) 667-7000
kys@lanl.gov

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