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The RNA accordion model for template positioning by telomerase RNA during telomeric DNA synthesis

Nature Structural & Molecular Biology volume 18, pages 1371–1375 (2011)Cite this article

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Abstract

Telomerase is a ribonucleoprotein (RNP) enzyme that maintains the ends of linear eukaryotic chromosomes and whose activation is a hallmark of 90% of all cancers. This RNP minimally contains a reverse transcriptase protein subunit (TERT) that catalyzes telomeric DNA synthesis and an RNA subunit (TER) that has templating, architectural and protein-scaffolding roles. Telomerase is unique among polymerases in that it synthesizes multiple copies of the template on the 3′ end of a primer following a single binding event, a process known as repeat addition processivity (RAP). Using biochemical assays and single-molecule Förster resonance energy transfer (smFRET) experiments on Tetrahymena thermophila telomerase, we now directly demonstrate that TER contributes to template positioning within the active site and to the template translocation required for RAP. We propose that the single-stranded RNA elements flanking the template act as a molecular accordion, undergoing reciprocal extension and compaction during telomerase translocation.

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Figure 1: The RNA accordion model for primer positioning and translocation.
Figure 2: Deletion mutants in the TRE have predictable primer usage profiles.
Figure 3: The accordion model predicts primer usage for TRE deletion mutants.
Figure 4: Single-molecule FRET measurements of RNA compression and extension at successive steps of telomere repeat synthesis.

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Acknowledgements

This work was partially supported by a Jane Coffin Childs Memorial Fund for Medical Research postdoctoral fellowship to A.J.B., US National Institutes of Health grant GM095850 to M.D.S. and US National Institutes of Health training grant T32 GM8646 to B.M.A.

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Author notes

  1. Andrea J Berman and Benjamin M Akiyama: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry and Biochemistry, Howard Hughes Medical Institute, University of Colorado, Boulder, Colorado, USA

    Andrea J Berman & Thomas R Cech

  2. Department of Molecular, Cell and Developmental Biology, University of California, Santa Cruz, California, USA

    Benjamin M Akiyama

  3. Department of Chemistry and Biochemistry, University of California, Santa Cruz, California, USA

    Michael D Stone

Authors
  1. Andrea J Berman
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  2. Benjamin M Akiyama
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Contributions

A.J.B. conducted all biochemical experiments; A.J.B. and T.R.C. designed and analyzed biochemical experiments. B.M.A. conducted smFRET experiments; B.M.A. and M.D.S. designed and analyzed smFRET experiments. All authors participated in writing the manuscript.

Corresponding author

Correspondence to Thomas R Cech.

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The authors declare no competing financial interests.

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Berman, A., Akiyama, B., Stone, M. et al. The RNA accordion model for template positioning by telomerase RNA during telomeric DNA synthesis. Nat Struct Mol Biol 18, 1371–1375 (2011). https://doi.org/10.1038/nsmb.2174

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