Structural and functional basis of mammalian microRNA biogenesis by Dicer

David Zapletal, Eliska Taborska, Josef Pasulka, Radek Malik, Karel Kubicek, Martina Zanova, Christian Much, Marek Sebesta, Valeria Buccheri, Filip Horvat, Irena Jenickova, Michaela Prochazkova, Jan Prochazka, Matyas Pinkas, Jiri Novacek, Diego F. Joseph, Radislav Sedlacek, Carrie Bernecky, Dónal O'Carroll, Richard Stefl*Petr Svoboda

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

MicroRNA (miRNA) and RNA interference (RNAi) pathways rely on small RNAs produced by Dicer endonucleases. Mammalian Dicer primarily supports the essential gene-regulating miRNA pathway, but how it is specifically adapted to miRNA biogenesis is unknown. We show that the adaptation entails a unique structural role of Dicer's DExD/H helicase domain. Although mice tolerate loss of its putative ATPase function, the complete absence of the domain is lethal because it assures high-fidelity miRNA biogenesis. Structures of murine Dicer•–miRNA precursor complexes revealed that the DExD/H domain has a helicase-unrelated structural function. It locks Dicer in a closed state, which facilitates miRNA precursor selection. Transition to a cleavage-competent open state is stimulated by Dicer-binding protein TARBP2. Absence of the DExD/H domain or its mutations unlocks the closed state, reduces substrate selectivity, and activates RNAi. Thus, the DExD/H domain structurally contributes to mammalian miRNA biogenesis and underlies mechanistical partitioning of miRNA and RNAi pathways.

Original languageEnglish
Pages (from-to)4064-4079.e13
Number of pages17
JournalMolecular Cell
Volume82
Issue number21
DOIs
Publication statusPublished - 3 Nov 2022

Keywords

  • cryo-EM
  • DExD
  • Dicer
  • dsRBD
  • dsRNA
  • helicase
  • miRNA
  • mirtron
  • PKR
  • RNAi
  • TARBP2

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