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Precursors of tRNAs are stabilized by methylguanosine cap structures

Abstract

Efficient maturation of transfer RNAs (tRNAs) is required for rapid cell growth. However, the precise timing of tRNA processing in coordination with the order of tRNA modifications has not been thoroughly elucidated. To analyze the modification status of tRNA precursors (pre-tRNAs) during maturation, we isolated pre-tRNAs at various stages from Saccharomyces cerevisiae and subjected them to MS analysis. We detected methylated guanosine cap structures at the 5′ termini of pre-tRNAs bearing 5′ leader sequences. These capped pre-tRNAs accumulated substantially after inhibition of RNase P activity. Upon depletion of the capping enzyme Ceg1p, the steady state level of capped pre-tRNA was markedly reduced. In addition, a population of capped pre-tRNAs accumulated in strains in which 5′ exonucleases were inhibited, indicating that the 5′ cap structures protect pre-tRNAs from 5′-exonucleolytic degradation during maturation.

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Figure 1: tRNA maturation in S. cerevisiae and accumulation of pre-tRNA in RNase P–repressed cells.
Figure 2: Methylguanosine cap structures found at 5′ termini of precursors from intronless tRNA genes in RNase P–repressed cells of S. cerevisiae.
Figure 3: Pre-tRNA capping of the intron-containing tRNAs in RNase P–repressed cells of S. cerevisiae.
Figure 4: Pre-tRNA capping in normally growing WT cells of S. cerevisiae.
Figure 5: Biosynthesis of pre-tRNA capping.
Figure 6: Pre-tRNA capping protects tRNA precursors from 5′-exonucleolytic degradation.

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Acknowledgements

We thank Y. Sakaguchi, K. Miyauchi, T. Suzuki and other members of T.S.'s laboratory for technical support and discussion. We also thank Y. Ohya (University of Tokyo), T. Yoshihisa (University of Hyogo) and M. Werner (Institute of Biology and Technology–Saclay) for providing materials. This work was supported by Grants-in-Aid for Young Scientists (B) (T.O.) and by Grants-in-Aid for Scientific Research on Priority Areas from the Ministry of Education, Science, Sports, and Culture of Japan (T.S.).

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T.O. and T.S. designed and performed the experiments and wrote the manuscript. T.S. supervised all the work.

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Correspondence to Tsutomu Suzuki.

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Supplementary Results, Supplementary Tables 1–4 and Supplementary Figures 1–13. (PDF 3165 kb)

Supplementary Data Set 1

RLE values of CAGE analysis in tRNA genes. (XLSX 210 kb)

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Ohira, T., Suzuki, T. Precursors of tRNAs are stabilized by methylguanosine cap structures. Nat Chem Biol 12, 648–655 (2016). https://doi.org/10.1038/nchembio.2117

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