Abstract
Eri1 is a 3′-to-5′ exoribonuclease conserved from fission yeast to humans. Here we show that Eri1 associates with ribosomes and ribosomal RNA (rRNA). Ribosomes from Eri1–deficient mice contain 5.8S rRNA that is aberrantly extended at its 3′ end, and Eri1, but not a catalytically inactive mutant, converts this abnormal 5.8S rRNA to the wild-type form in vitro and in cells. In human and murine cells, Eri1 localizes to the cytoplasm and nucleus, with enrichment in the nucleolus, the site of preribosome biogenesis. RNA binding residues in the Eri1 SAP and linker domains promote stable association with rRNA and thereby facilitate 5.8S rRNA 3′ end processing. Taken together, our findings indicate that Eri1 catalyzes the final trimming step in 5.8S rRNA processing, functionally and spatially connecting this regulator of RNAi with the basal translation machinery.
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Acknowledgements
We wish to thank W. Hammerschmidt, Helmholtz Center Munich, for the gift of the SV40 large T hygro retrovirus, D. Eick, Helmholtz Center Munich, for the gift of anti–PES-1 antibody and F. Alt, Immune Disease Institute, Boston, for recombinant Cre adenovirus. We thank J. Asara for assistance with MS and data analysis, E. Yanni and C. Gelinas for technical assistance and A. Whynot, T.A. Rapoport and P. Sorger for help and equipment for gradient fractionation. We also thank H. Gabel and G. Ruvkun for communicating unpublished data, the laboratories of D. Eick, M. Meisterernst and I. Jeremias for helpful discussion, and D. Eick and D. Moazed for comments on the manuscript. This work was supported by US National Institutes of Health (NIH) grants AI44432 and AI70788 to A.R., a Cancer Research Institute fellowship and Deutsche Forschungsgemeinschaft grant HE 3359/2 to V.H., and a Burroughs Wellcome Fund Career Award in the Biomedical Sciences to K.M.A. K.M.A. is a Special Fellow of The Leukemia and Lymphoma Society. W.A.P. is a National Defense Science and Engineering Graduate fellow. M.T. is supported by the Harvard Stem Cell Institute and is a Predoctoral Fellow of the Ryan Foundation.
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K.M.A. and V.H. initiated the project, planned and supervised the experiments, performed some experiments, and wrote the manuscript. A.R. provided advice and support and edited the manuscript. W.A.P., N.R. and A.D.L. established tools and assays and performed and analyzed experiments with technical assistance and advice from E.G., C.W., L.C.S., N.P., E.D.L., M.T., J.W.E. and Y.S. The monoclonal anti-Eri1 antibody was generated by E.K.
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Ansel, K., Pastor, W., Rath, N. et al. Mouse Eri1 interacts with the ribosome and catalyzes 5.8S rRNA processing. Nat Struct Mol Biol 15, 523–530 (2008). https://doi.org/10.1038/nsmb.1417
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DOI: https://doi.org/10.1038/nsmb.1417
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