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An mRNA-rRNA base-pairing mechanism for translation initiation in eukaryotes

Nature Structural & Molecular Biology volume 13, pages 30–34 (2006)Cite this article

Abstract

Base-pairing of messenger RNA to ribosomal RNA is a mechanism of translation initiation in prokaryotes. Although analogous base-pairing has been suggested to affect the translation of various eukaryotic mRNAs, direct evidence has been lacking. To test such base-pairing, we developed a yeast system that uses ribosomes containing a mouse-yeast hybrid 18S rRNA. Using this system, we demonstrate that a 9-nucleotide element found in the mouse Gtx homeodomain mRNA facilitates translation initiation by base-pairing to 18S rRNA. Various point mutations in the Gtx element and in either the hybrid or wild-type yeast 18S rRNAs confirmed the requirement for an intact complementary match. The presence of the Gtx element in various mRNAs suggests that this element affects the translation of groups of mRNAs. We discuss the possibility that other mRNA elements affect translation by base-pairing to different sites in the 18S rRNA.

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Figure 1: The Gtx translational enhancer element is active in mammalian cells but inactive in yeast.
Figure 2: The Gtx TEE can function in yeast that express hybrid ribosomes containing mouse 18S rRNA.
Figure 3: Activity of the Gtx TEE requires a complementary match to mouse 18S rRNA.
Figure 4: The Gtx translational enhancer element can function in yeast when three nucleotides of the yeast 18S rRNA are mutated.

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Acknowledgements

We thank M. Nomura at the University of California, Irvine, California, USA for providing us with yeast strain NOY908 and plasmids pNOY353 and pNOY373. We also thank L Burman for excellent technical assistance and G.M. Edelman, B.A. Cunningham and J.A. Gally for valuable comments and critical reading of the manuscript. Funding was provided by the US National Institutes of Health (grant GM61725) and the G. Harold and Leila Y. Mathers Charitable Foundation to V.P.M, and by the Skaggs Institute for Chemical Biology to J.D., S.A.C. and W.Z.

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  1. Department of Neurobiology, The Scripps Research Institute, and The Skaggs Institute for Chemical Biology, 10550 North Torrey Pines Road, La Jolla, 92037, California, USA

    John Dresios, Stephen A Chappell, Wei Zhou & Vincent P Mauro

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  1. John Dresios
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Correspondence to Vincent P Mauro.

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Supplementary information

Supplementary Fig. 1

Expression of hybrid rRNAs in yeast. (PDF 140 kb)

Supplementary Fig. 2

Quantitative northern blot analysis. (PDF 112 kb)

Supplementary Table 1

Expression vectors (PDF 78 kb)

Supplementary Table 2

Yeast strains and plasmids (PDF 89 kb)

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Dresios, J., Chappell, S., Zhou, W. et al. An mRNA-rRNA base-pairing mechanism for translation initiation in eukaryotes. Nat Struct Mol Biol 13, 30–34 (2006). https://doi.org/10.1038/nsmb1031

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