Regulation of RUVBL1-RUVBL2 AAA-ATPases by the nonsense-mediated mRNA decay factor DHX34, as evidenced by Cryo-EM
- Spanish National Cancer Research Center, CNIO, Spain
- MRC Human Genetics Unit, Institute of Genetics and Molecular Medicine, University of Edinburgh, United Kingdom
Abstract
Nonsense-mediated mRNA decay (NMD) is a surveillance pathway that degrades aberrant mRNAs and also regulates the expression of a wide range of physiological transcripts. RUVBL1 and RUVBL2 AAA-ATPases form an hetero-hexameric ring that is part of several macromolecular complexes such as INO80, SWR1 and R2TP. Interestingly, RUVBL1-RUVBL2 ATPase activity is required for NMD activation by an unknown mechanism. Here, we show that DHX34, an RNA helicase regulating NMD initiation, directly interacts with RUVBL1-RUVBL2 in vitro and in cells. Cryo-EM reveals that DHX34 induces extensive changes in the N-termini of every RUVBL2 subunit in the complex, stabilizing a conformation that does not bind nucleotide and thereby down-regulates ATP hydrolysis of the complex. Using ATPase-deficient mutants, we find that DHX34 acts exclusively on the RUVBL2 subunits. We propose a model, where DHX34 acts to couple RUVBL1-RUVBL2 ATPase activity to the assembly of factors required to initiate the NMD response.
Data availability
The cryo-EM maps of the RUVBL1-RUVBL2-DHX34 complex and the RUVBL1-RUVBL2 ring have been deposited in the EM database with accession codes EMD-11788 and EMD-11789 respectively. The structure of RUVBL1-RUVBL2 heterohexameric ring after binding of RNA helicase DHX34 has been deposited as PDB ID 7AHO.
Article and author information
Author details
Funding
Spanish Ministry of Science and Innovation (SAF2017-82632-P)
- Andrés López-Perrote
- Carlos F Rodríguez
- Marina Serna
- Oscar Llorca
Autonomous Government of Madrid (Y2018/BIO4747)
- Ana González-Corpas
- Oscar Llorca
Autonomous Government of Madrid (P2018/NMT4443)
- Ana González-Corpas
- Oscar Llorca
Core funding to the MRC Human Genetics Unit
- Javier F Caceres
Spanish Ministry of Science and Innovation (BES-2015-071348)
- Carlos F Rodríguez
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Andreas Martin, University of California, Berkeley, United States
Version history
- Received: September 12, 2020
- Accepted: November 12, 2020
- Accepted Manuscript published: November 18, 2020 (version 1)
- Version of Record published: December 1, 2020 (version 2)
Copyright
© 2020, López-Perrote et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Regulation of RUVBL1-RUVBL2 AAA-ATPases by the nonsense-mediated mRNA decay factor DHX34, as evidenced by Cryo-EM
eLife 9:e63042.
https://doi.org/10.7554/eLife.63042
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