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How mutations in tRNA distant from the anticodon affect the fidelity of decoding

Schmeing, T. Martin and Voorhees, Rebecca M. and Kelley, Ann C. and Ramakrishnan, V. (2011) How mutations in tRNA distant from the anticodon affect the fidelity of decoding. Nature Structural and Molecular Biology, 18 (4). pp. 432-436. ISSN 1545-9993. PMCID PMC3072312. https://resolver.caltech.edu/CaltechAUTHORS:20180119-163125316

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Abstract

The ribosome converts genetic information into protein by selecting aminoacyl tRNAs whose anticodons base-pair to an mRNA codon. Mutations in the tRNA body can perturb this process and affect fidelity. The Hirsh suppressor is a well-studied tRNA^(Trp) harboring a G24A mutation that allows readthrough of UGA stop codons. Here we present crystal structures of the 70S ribosome complexed with EF-Tu and aminoacyl tRNA (native tRNA^(Trp), G24A tRNA^(Trp) or the miscoding A9C tRNA^(Trp)) bound to cognate UGG or near-cognate UGA codons, determined at 3.2-Å resolution. The A9C and G24A mutations lead to miscoding by facilitating the distortion of tRNA required for decoding. A9C accomplishes this by increasing tRNA flexibility, whereas G24A allows the formation of an additional hydrogen bond that stabilizes the distortion. Our results also suggest that each native tRNA will adopt a unique conformation when delivered to the ribosome that allows accurate decoding.


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https://doi.org/10.1038/nsmb.2003DOIArticle
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https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3072312/PubMed CentralArticle
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ORCID:
AuthorORCID
Voorhees, Rebecca M.0000-0003-1640-2293
Additional Information:© 2011 Macmillan Publishers Limited. Received 19 October 2010; accepted 8 December 2010; published online 6 March 2011; doi:10.1038/nsmb.2003 We thank R. Green and R. Ortiz-Meoz, Johns Hopkins University, for plasmids and bacterial strains for production of mutant tRNA^(Trp), A. McCarthy and S. Brockhauser at ESRF ID14.4 for facilitating data collection, O. Uhlenbeck for helpful discussion and F. Murphy for scripting. This work was supported by the Medical Research Council, the Wellcome Trust, the Agouron Institute and the Louis-Jeantet Foundation. R.M.V. received support from the Gates-Cambridge scholarship. T.M.S. received support from the Human Frontier Science Program and Emmanuel College. Author Contributions: T.M.S. and R.M.V. designed the experiments, prepared and crystallized the ribosomal complexes, collected and processed X-ray crystallography data, determined, refined and analyzed the structures, and prepared the manuscript and figures. A.C.K. purified macromolecular components, prepared and crystallized ribosomal complexes and aided with collection of X-ray crystallography data. V.R. suggested the study and provided intellectual input and supervision. Accession codes. Protein Data Bank: 2Y0U, 2Y0V, 2Y0W, 2Y0X for A9C; 2Y0Y, 2Y0Z, 2Y12, 2Y13 for HS; 2Y14, 2Y15, 2Y16, 2Y17 for HT; and 2Y18, 2Y19, 2Y10, 2Y11 for TT. Competing interests: V.R. is on the Senior Advisory Board of Rib-X Pharmaceuticals and both T.M.S. and V.R. hold stock options.
Funders:
Funding AgencyGrant Number
Medical Research Council (UK)UNSPECIFIED
Wellcome TrustUNSPECIFIED
Agouron InstituteUNSPECIFIED
Louis-Jeantet FoundationUNSPECIFIED
Gates-Cambridge ScholarshipUNSPECIFIED
Human Frontier Science ProgramUNSPECIFIED
Emmanuel CollegeUNSPECIFIED
Issue or Number:4
PubMed Central ID:PMC3072312
Record Number:CaltechAUTHORS:20180119-163125316
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20180119-163125316
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:84436
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:22 Jan 2018 15:57
Last Modified:01 Nov 2019 22:16

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