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Regulation of RUVBL1-RUVBL2 AAA-ATPases by the nonsense-mediated mRNA decay factor DHX34, as evidenced by Cryo-EM

Andres López-Perrote1, Nele Hug2, Ana González-Corpas1

  • 1Structural Biology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain.

Elife
|November 18, 2020
PubMed

Insights

The RNA helicase DHX34 regulates nonsense-mediated mRNA decay (NMD) by interacting with RUVBL1-RUVBL2 ATPases. DHX34 stabilizes a conformation that down-regulates ATPase activity, impacting NMD initiation.

Area of Science:

  • Molecular Biology
  • Cellular Biology
  • Biochemistry

Background:

  • Nonsense-mediated mRNA decay (NMD) is a crucial surveillance pathway for degrading aberrant mRNAs and regulating gene expression.
  • RUVBL1 and RUVBL2 form a hetero-hexameric ring involved in various macromolecular complexes, with their ATPase activity essential for NMD activation.

Purpose of the Study:

  • To elucidate the mechanism by which RUVBL1-RUVBL2 ATPase activity is regulated for NMD activation.
  • To investigate the interaction between the RNA helicase DHX34 and the RUVBL1-RUVBL2 complex in the context of NMD.

Main Methods:

  • In vitro and cellular interaction studies between DHX34 and RUVBL1-RUVBL2.
  • Cryo-electron microscopy (Cryo-EM) to determine the structural basis of the interaction.
  • Biochemical assays using ATPase-deficient mutants of RUVBL1-RUVBL2.

Main Results:

  • DHX34 directly interacts with the RUVBL1-RUVBL2 complex.
  • Cryo-EM revealed DHX34 induces conformational changes in RUVBL2 N-termini, inhibiting nucleotide binding and ATP hydrolysis.
  • DHX34's effect is specific to the RUVBL2 subunits, as shown by ATPase-deficient mutants.

Conclusions:

  • DHX34 acts as a regulator of RUVBL1-RUVBL2 ATPase activity, specifically targeting RUVBL2 subunits.
  • A model is proposed where DHX34 couples RUVBL1-RUVBL2 ATPase activity to the assembly of NMD-initiating factors.
Keywords:
AAA+ ATPasesDHX34RUVBL1-RUVBL2humanmolecular biophysicsnonsense mediated mRNA decaystructural biology

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