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Single-molecule FRET uncovers hidden conformations and dynamics of human Argonaute 2

Author

Listed:
  • Sarah Willkomm

    (University of Regensburg)

  • Leonhard Jakob

    (University of Regensburg
    University of Regensburg)

  • Kevin Kramm

    (University of Regensburg)

  • Veronika Graus

    (University of Regensburg)

  • Julia Neumeier

    (University of Regensburg)

  • Gunter Meister

    (University of Regensburg)

  • Dina Grohmann

    (University of Regensburg
    University of Regensburg)

Abstract

Human Argonaute 2 (hAgo2) constitutes the functional core of the RNA interference pathway. Guide RNAs direct hAgo2 to target mRNAs, which ultimately leads to hAgo2-mediated mRNA degradation or translational inhibition. Here, we combine site-specifically labeled hAgo2 with time-resolved single-molecule FRET measurements to monitor conformational states and dynamics of hAgo2 and hAgo2-RNA complexes in solution that remained elusive so far. We observe dynamic anchoring and release of the guide’s 3’-end from the PAZ domain during the stepwise target loading process even with a fully complementary target. We find differences in structure and dynamic behavior between partially and fully paired canonical hAgo2-guide/target complexes and the miRNA processing complex formed by hAgo2 and pre-miRNA451. Furthermore, we detect a hitherto unknown conformation of hAgo2-guide/target complexes that poises them for target-directed miRNA degradation. Taken together, our results show how the conformational flexibility of hAgo2-RNA complexes determines function and the fate of the ribonucleoprotein particle.

Suggested Citation

  • Sarah Willkomm & Leonhard Jakob & Kevin Kramm & Veronika Graus & Julia Neumeier & Gunter Meister & Dina Grohmann, 2022. "Single-molecule FRET uncovers hidden conformations and dynamics of human Argonaute 2," Nature Communications, Nature, vol. 13(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-31480-4
    DOI: 10.1038/s41467-022-31480-4
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    References listed on IDEAS

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    1. Yanli Wang & Gang Sheng & Stefan Juranek & Thomas Tuschl & Dinshaw J. Patel, 2008. "Structure of the guide-strand-containing argonaute silencing complex," Nature, Nature, vol. 456(7219), pages 209-213, November.
    2. Yanli Wang & Stefan Juranek & Haitao Li & Gang Sheng & Greg S. Wardle & Thomas Tuschl & Dinshaw J. Patel, 2009. "Nucleation, propagation and cleavage of target RNAs in Ago silencing complexes," Nature, Nature, vol. 461(7265), pages 754-761, October.
    3. Yanli Wang & Stefan Juranek & Haitao Li & Gang Sheng & Thomas Tuschl & Dinshaw J. Patel, 2008. "Structure of an argonaute silencing complex with a seed-containing guide DNA and target RNA duplex," Nature, Nature, vol. 456(7224), pages 921-926, December.
    4. Michael J. Moore & Troels K. H. Scheel & Joseph M. Luna & Christopher Y. Park & John J. Fak & Eiko Nishiuchi & Charles M. Rice & Robert B. Darnell, 2015. "miRNA–target chimeras reveal miRNA 3′-end pairing as a major determinant of Argonaute target specificity," Nature Communications, Nature, vol. 6(1), pages 1-17, December.
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