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Telomerase RNA structural heterogeneity in living human cells detected by DMS-MaPseq

Author

Listed:
  • Nicholas M. Forino

    (University of California)

  • Jia Zheng Woo

    (Harvard Medical School)

  • Arthur J. Zaug

    (University of Colorado
    University of Colorado)

  • Arcelia Gonzalez Jimenez

    (University of California)

  • Eva Edelson

    (University of California)

  • Thomas R. Cech

    (University of Colorado
    University of Colorado)

  • Silvi Rouskin

    (Harvard Medical School)

  • Michael D. Stone

    (University of California)

Abstract

Biogenesis of human telomerase requires its RNA subunit (hTR) to fold into a multi-domain architecture that includes the template-pseudoknot (t/PK) and the three-way junction (CR4/5). These hTR domains bind the telomerase reverse transcriptase (hTERT) protein and are essential for telomerase activity. Here, we probe hTR structure in living cells using dimethyl sulfate mutational profiling with sequencing (DMS-MaPseq) and ensemble deconvolution analysis. Approximately 15% of the steady state population of hTR has a CR4/5 conformation lacking features required for hTERT binding. The proportion of hTR CR4/5 folded into the primary functional conformation is independent of hTERT expression levels. Mutations that stabilize the alternative CR4/5 conformation are detrimental to telomerase assembly and activity. Moreover, the alternative CR4/5 conformation is not found in purified telomerase RNP complexes, supporting the hypothesis that only the primary CR4/5 conformer is active. We propose that this misfolded portion of the cellular hTR pool is either slowly refolded or degraded, suggesting that kinetic RNA folding traps studied in vitro may also hinder ribonucleoprotein assembly in vivo.

Suggested Citation

  • Nicholas M. Forino & Jia Zheng Woo & Arthur J. Zaug & Arcelia Gonzalez Jimenez & Eva Edelson & Thomas R. Cech & Silvi Rouskin & Michael D. Stone, 2025. "Telomerase RNA structural heterogeneity in living human cells detected by DMS-MaPseq," Nature Communications, Nature, vol. 16(1), pages 1-15, December.
    • Handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-56149-6
    DOI: 10.1038/s41467-025-56149-6
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    References listed on IDEAS

    as
    1. Baocheng Liu & Yao He & Yaqiang Wang & He Song & Z. Hong Zhou & Juli Feigon, 2022. "Structure of active human telomerase with telomere shelterin protein TPP1," Nature, Nature, vol. 604(7906), pages 578-583, April.
    2. Michael D. Stone & Mariana Mihalusova & Catherine M. O’Connor & Ramadevi Prathapam & Kathleen Collins & Xiaowei Zhuang, 2007. "Stepwise protein-mediated RNA folding directs assembly of telomerase ribonucleoprotein," Nature, Nature, vol. 446(7134), pages 458-461, March.
    3. Chi-Kang Tseng & Hui-Fang Wang & Morgan R. Schroeder & Peter Baumann, 2018. "The H/ACA complex disrupts triplex in hTR precursor to permit processing by RRP6 and PARN," Nature Communications, Nature, vol. 9(1), pages 1-12, December.
    4. Feng Wang & Elaine R. Podell & Arthur J. Zaug & Yuting Yang & Paul Baciu & Thomas R. Cech & Ming Lei, 2007. "The POT1–TPP1 telomere complex is a telomerase processivity factor," Nature, Nature, vol. 445(7127), pages 506-510, February.
    5. Tom Vulliamy & Anna Marrone & Frederick Goldman & Andrew Dearlove & Monica Bessler & Philip J. Mason & Inderjeet Dokal, 2001. "The RNA component of telomerase is mutated in autosomal dominant dyskeratosis congenita," Nature, Nature, vol. 413(6854), pages 432-435, September.
    6. Yao He & He Song & Henry Chan & Baocheng Liu & Yaqiang Wang & Lukas Sušac & Z. Hong Zhou & Juli Feigon, 2022. "Structure of Tetrahymena telomerase-bound CST with polymerase α-primase," Nature, Nature, vol. 608(7924), pages 813-818, August.
    7. George E. Ghanim & Adam J. Fountain & Anne-Marie M. Roon & Ramya Rangan & Rhiju Das & Kathleen Collins & Thi Hoang Duong Nguyen, 2021. "Structure of human telomerase holoenzyme with bound telomeric DNA," Nature, Nature, vol. 593(7859), pages 449-453, May.
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