Abstract
Although the catalytic subunit of the Schizosaccharomyces pombe telomerase holoenzyme was identified over ten years ago, the unusual heterogeneity of its telomeric DNA made it difficult to identify its RNA component. We used a new two-step immunoprecipitation and reverse transcription–PCR technique to identify the S. pombe telomerase RNA, which we call TER1. TER1 RNA was 1,213 nucleotides long, similar in size to the Saccharomyces cerevisiae telomerase RNA, TLC1. TER1 RNA associated in vivo with the two known subunits of the S. pombe telomerase holoenzyme, Est1p and Trt1p, and neither association was dependent on the other holoenzyme component. We present a model to explain how telomerase introduces heterogeneity into S. pombe telomeres. The technique used here to identify TER1 should be generally applicable to other model organisms.
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Acknowledgements
We thank P. Baumann for sharing results before publication and J.B. Boulé, M. Mateyak, J. Phillips, S. Pinter, M. Sabourin, C. Tuzon and Y. Wu for critical reading of the manuscript. We thank O. Troyanskya and C. Huttenhower for computational assistance, R. Allshire (Wellcome Trust Centre for Cell Biology), P. Baumann (Stowers Institute for Medical Research), H. Lieberman (Columbia University) and M. Sipiczki (University of Debrecen) for strains, and K. Miller in J. Cooper's lab, C. Tuzon and J. Bruzik for protocols. This work was supported by US National Institutes of Health grants GM43265 and R37 GM26938 and by a US National Research Service Award to C.J.W.
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Webb, C., Zakian, V. Identification and characterization of the Schizosaccharomyces pombe TER1 telomerase RNA. Nat Struct Mol Biol 15, 34–42 (2008). https://doi.org/10.1038/nsmb1354
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DOI: https://doi.org/10.1038/nsmb1354
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