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Spliceosomal cleavage generates the 3′ end of telomerase RNA

Nature volume 456, pages 910–914 (2008)Cite this article

Abstract

Telomeres cap the ends of chromosomes and provide a means to complete replication. The DNA portion of telomeres is synthesized by the enzyme telomerase using part of an RNA subunit as a template for reverse transcription. How the mature 3′ end of telomerase RNA is generated has so far remained elusive. Here we show that in Schizosaccharomyces pombe telomerase RNA transcripts must be processed to generate functional telomerase. Characterization of the maturation pathway uncovered an unexpected role for the spliceosome, which normally catalyses splicing of pre-messenger RNA. The first spliceosomal cleavage reaction generates the mature 3′ end of telomerase RNA (TER1, the functional RNA encoded by the ter1+ gene), releasing the active form of the RNA without exon ligation. Blocking the first step or permitting completion of splicing generates inactive forms of TER1 and causes progressive telomere shortening. We establish that 3′ end processing of TER1 is critical for telomerase function and describe a previously unknown mechanism for RNA maturation that uses the ability of the spliceosome to mediate site-specific cleavage.

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Figure 1: An intron in TER1 is essential for RNA maturation.
Figure 2: Splicing and the Sm binding site.
Figure 3: A direct role for the U1 snRNA in TER1 maturation.
Figure 4: The first and second transesterification reactions have opposing effects on TER1 maturation.
Figure 5: Requirements for 3′ end processing by the spliceosome.

References

  1. Hug, N. & Lingner, J. Telomere length homeostasis. Chromosoma 115, 413–425 (2006)

    Article  CAS  Google Scholar 

  2. Stewart, S. A. & Weinberg, R. A. Telomeres: cancer to human aging. Annu. Rev. Cell Dev. Biol. 22, 531–557 (2006)

    Article  CAS  Google Scholar 

  3. Mitchell, J. R., Wood, E. & Collins, K. A telomerase component is defective in the human disease dyskeratosis congenita. Nature 402, 551–555 (1999)

    Article  ADS  CAS  Google Scholar 

  4. Vulliamy, T. et al. The RNA component of telomerase is mutated in autosomal dominant dyskeratosis congenita. Nature 413, 432–435 (2001)

    Article  ADS  CAS  Google Scholar 

  5. Vulliamy, T., Marrone, A., Dokal, I. & Mason, P. J. Association between aplastic anaemia and mutations in telomerase RNA. Lancet 359, 2168–2170 (2002)

    Article  CAS  Google Scholar 

  6. Armanios, M. Y. et al. Telomerase mutations in families with idiopathic pulmonary fibrosis. N. Engl. J. Med. 356, 1317–1326 (2007)

    Article  CAS  Google Scholar 

  7. Yamaguchi, H. et al. Mutations in TERT, the gene for telomerase reverse transcriptase, in aplastic anemia. N. Engl. J. Med. 352, 1413–1424 (2005)

    Article  CAS  Google Scholar 

  8. Mitchell, J. R., Wood, E. & Collins, K. A telomerase component is defective in the human disease dyskeratosis congenita. Nature 402, 551–555 (1999)

    Article  ADS  CAS  Google Scholar 

  9. Cohen, S. B. et al. Protein composition of catalytically active human telomerase from immortal cells. Science 315, 1850–1853 (2007)

    Article  ADS  CAS  Google Scholar 

  10. Lingner, J. et al. Reverse transcriptase motifs in the catalytic subunit of telomerase. Science 276, 561–567 (1997)

    Article  CAS  Google Scholar 

  11. Gillis, A. J., Schuller, A. P. & Skordalakes, E. Structure of the Tribolium castaneum telomerase catalytic subunit TERT. Nature 455, 633–637 (2008)

    Article  ADS  CAS  Google Scholar 

  12. Leonardi, J., Box, J. A., Bunch, J. T. & Baumann, P. TER1, the RNA subunit of fission yeast telomerase. Nature Struct. Mol. Biol. 15, 26–33 (2008)

    Article  CAS  Google Scholar 

  13. Webb, C. J. & Zakian, V. A. Identification and characterization of the Schizosaccharomyces pombe TER1 telomerase RNA. Nature Struct. Mol. Biol. 15, 34–42 (2008)

    Article  CAS  Google Scholar 

  14. Baumann, P. & Cech, T. R. Pot1, the putative telomere end-binding protein in fission yeast and humans. Science 292, 1171–1175 (2001)

    Article  ADS  CAS  Google Scholar 

  15. Chikashige, Y. & Hiraoka, Y. Telomere binding of the Rap1 protein is required for meiosis in fission yeast. Curr. Biol. 11, 1618–1623 (2001)

    Article  CAS  Google Scholar 

  16. Cooper, J. P., Nimmo, E. R., Allshire, R. C. & Cech, T. R. Regulation of telomere length and function by a Myb-domain protein in fission yeast. Nature 385, 744–747 (1997)

    Article  ADS  CAS  Google Scholar 

  17. Kanoh, J. & Ishikawa, F. spRap1 and spRif1, recruited to telomeres by Taz1, are essential for telomere function in fission yeast. Curr. Biol. 11, 1624–1630 (2001)

    Article  CAS  Google Scholar 

  18. Miyoshi, T., Kanoh, J., Saito, M. & Ishikawa, F. Fission yeast Pot1–Tpp1 protects telomeres and regulates telomere length. Science 320, 1341–1344 (2008)

    Article  ADS  CAS  Google Scholar 

  19. Chapon, C., Cech, T. R. & Zaug, A. J. Polyadenylation of telomerase RNA in budding yeast. RNA 3, 1337–1351 (1997)

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Fu, D. & Collins, K. Distinct biogenesis pathways for human telomerase RNA and H/ACA small nucleolar RNAs. Mol. Cell 11, 1361–1372 (2003)

    Article  CAS  Google Scholar 

  21. Zhang, M. Q. & Marr, T. G. Fission yeast gene structure and recognition. Nucleic Acids Res. 22, 1750–1759 (1994)

    Article  CAS  Google Scholar 

  22. Seto, A. G., Zaug, A. J., Sobel, S. G., Wolin, S. L. & Cech, T. R. Saccharomyces cerevisiae telomerase is an Sm small nuclear ribonucleoprotein particle. Nature 401, 177–180 (1999)

    Article  ADS  CAS  Google Scholar 

  23. Seraphin, B., Kretzner, L. & Rosbash, M. A. U1 snRNA:pre-mRNA base pairing interaction is required early in yeast spliceosome assembly but does not uniquely define the 5′ cleavage site. EMBO J. 7, 2533–2538 (1988)

    Article  CAS  Google Scholar 

  24. Siliciano, P. G. & Guthrie, C. 5′ splice site selection in yeast: genetic alterations in base-pairing with U1 reveal additional requirements. Genes Dev. 2, 1258–1267 (1988)

    Article  CAS  Google Scholar 

  25. Zhuang, Y. & Weiner, A. M. A compensatory base change in U1 snRNA suppresses a 5′ splice site mutation. Cell 46, 827–835 (1986)

    Article  CAS  Google Scholar 

  26. Alvarez, C. J., Romfo, C. M., Vanhoy, R. W., Porter, G. L. & Wise, J. A. Mutational analysis of U1 function in Schizosaccharomyces pombe: pre-mRNAs differ in the extent and nature of their requirements for this snRNA in vivo . RNA 2, 404–418 (1996)

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Seraphin, B. & Rosbash, M. Mutational analysis of the interactions between U1 small nuclear RNA and pre-mRNA of yeast. Gene 82, 145–151 (1989)

    Article  CAS  Google Scholar 

  28. Domdey, H. et al. Lariat structures are in vivo intermediates in yeast pre-mRNA splicing. Cell 39, 611–621 (1984)

    Article  CAS  Google Scholar 

  29. Romfo, C. M. & Wise, J. A. Both the polypyrimidine tract and the 3′ splice site function prior to the first step of splicing in fission yeast. Nucleic Acids Res. 25, 4658–4665 (1997)

    Article  CAS  Google Scholar 

  30. Bunch, J. T., Bae, N. S., Leonardi, J. & Baumann, P. Distinct requirements for Pot1 in limiting telomere length and maintaining chromosome stability. Mol. Cell. Biol. 25, 5567–5578 (2005)

    Article  CAS  Google Scholar 

  31. Box, J. A., Bunch, J. T., Zappulla, D. C., Glynn, E. F. & Baumann, P. A flexible template boundary element in the RNA subunit of fission yeast telomerase. J. Biol. Chem. 283, 24224–24233 (2008)

    Article  CAS  Google Scholar 

  32. Burge, C. B., Tuschl, T. H. & Sharp, P. A. in The RNA World (eds Gesteland, R. F., Cech, T. R. & Atkins, J. F.) 525–560 (Cold Spring Harbor Laboratory Press, 1999)

    Google Scholar 

  33. Forsburg, S. L. Comparison of Schizosaccharomyces pombe expression systems. Nucleic Acids Res. 21, 2955–2956 (1993)

    Article  CAS  Google Scholar 

  34. Church, G. M. & Gilbert, W. Genomic sequencing. Proc. Natl Acad. Sci. USA 81, 1991–1995 (1984)

    Article  ADS  CAS  Google Scholar 

  35. Li, S. G., Zhou, H., Luo, Y. P., Zhang, P. & Qu, L. H. Identification and functional analysis of 20 Box H/ACA small nucleolar RNAs (snoRNAs) from Schizosaccharomyces pombe . J. Biol. Chem. 280, 16446–16455 (2005)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors thank H. Yang and the other members of the Baumann laboratory for help and discussions, Y. Tzfati and L. Tomaska for sharing results before publication, and A. Berglund, M. Blanchette, R. Conaway and T. Cech for discussions and comments on the manuscript. We also thank the Molecular Biology Core Facility for site-directed mutagenesis and sequencing, M. Gogol and R. Voelker for computational analysis, and D. Baumann and R. Helston for proofreading of the manuscript. This work was funded by the Stowers Institute for Medical Research and a Pew Scholars in the Biomedical Sciences Award to P.B.

Author Contributions P.B. made the initial observations, oversaw the project and designed the experiments. J.T.B. and P.B. developed protocols for RNA isolation, northern blotting and primer extension analysis. J.A.B. contributed plasmids and strains and performed telomere length analysis and RT–PCR assays. J.T.B. conducted northern blotting and primer extension analysis. W.T. characterized the TER1-Sm1 mutant. All authors contributed to data analysis, and P.B. wrote the manuscript.

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Author notes

  1. Jessica A. Box and Jeremy T. Bunch: These authors contributed equally to this work.

Authors and Affiliations

  1. Stowers Institute for Medical Research, Kansas City, Missouri 64110, USA ,

    Jessica A. Box, Jeremy T. Bunch, Wen Tang & Peter Baumann

  2. Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas 66160, USA,

    Wen Tang & Peter Baumann

Authors
  1. Jessica A. Box
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Corresponding author

Correspondence to Peter Baumann.

Supplementary information

Supplementary Information

This file contais Supplementary Figures 1-6 illustrating the 5′ splice site consensus in S. pombe, the effects of mutations in TER1 on splicing, the mapping of the branch point by primer extension and TER1 processing at various levels of expression. A detailed schematic of the heterologous intron and mutations therein is also provided. (PDF 1177 kb)

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Box, J., Bunch, J., Tang, W. et al. Spliceosomal cleavage generates the 3′ end of telomerase RNA. Nature 456, 910–914 (2008). https://doi.org/10.1038/nature07584

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