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The central core region of yeast ribosomal protein L11 is important for subunit joining and translational fidelity

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Abstract

Yeast ribosomal protein L11 is positioned at the intersubunit cleft of the large subunit central protuberance, forming an intersubunit bridge with the small subunit protein S18. Mutants were engineered in the central core region of L11 which interacts with Helix 84 of the 25S rRNA. Numerous mutants in this region conferred 60S subunit biogenesis defects. Specifically, many mutations of F96 and the A66D mutant promoted formation of halfmers as assayed by sucrose density ultracentrifugation. Halfmer formation was not due to deficiency in 60S subunit production, suggesting that the mutants affected subunit-joining. Chemical modification analyses indicated that the A66D mutant, but not the F96 mutants, promoted changes in 25S rRNA structure, suggesting at least two modalities for subunit joining defects. 25S rRNA structural changes were located both adjacent to A66D (in H84), and more distant (in H96-7). While none of the mutants significantly affected ribosome/tRNA binding constants, they did have strong effects on cellular growth at both high and low temperatures, in the presence of translational inhibitors, and promoted changes in translational fidelity. Two distinct mechanisms are proposed by which L11 mutants may affect subunit joining, and identification of the amino acids associated with each of these processes are presented. These findings may have implications for our understanding of multifaceted diseases such as Diamond–Blackfan anemia which have been linked in part with mutations in L11.

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Acknowledgments

We would like to thank Dr. Arturas Meskauskas, Dr. Karen Jack, Hamid-Reza Shahshahan, Ashton Trey Belew, Sharmishtha Musalgaonkar, and the rest of our laboratory for their help and support. We thank Dr. Pamela Silver for providing us with L11 yeast strain PSY2088. This work was supported by a grant to JDD from the National Institutes of Health (R01 GM058859).

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The authors declare they have no conflict of interest.

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  1. Department of Cell Biology and Molecular Genetics, University of Maryland, Microbiology Building Room 2135, College Park, MD, 20742, USA

    Michael H. J. Rhodin, Rasa Rakauskaitė & Jonathan D. Dinman

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  1. Michael H. J. Rhodin
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  2. Rasa Rakauskaitė
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  3. Jonathan D. Dinman
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Correspondence to Jonathan D. Dinman.

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Communicated by T. Ito.

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Rhodin, M.H.J., Rakauskaitė, R. & Dinman, J.D. The central core region of yeast ribosomal protein L11 is important for subunit joining and translational fidelity. Mol Genet Genomics 285, 505–516 (2011). https://doi.org/10.1007/s00438-011-0623-2

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  • DOI: https://doi.org/10.1007/s00438-011-0623-2

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