High-resolution structures of the actomyosin-V complex in three nucleotide states provide insights into the force generation mechanism
Abstract
The molecular motor myosin undergoes a series of major structural transitions during its force-producing motor cycle. The underlying mechanism and its coupling to ATP hydrolysis and actin binding is only partially understood, mostly due to sparse structural data on actin-bound states of myosin. Here, we report 26 high-resolution cryo-EM structures of the actomyosin-V complex in the strong-ADP, rigor, and a previously unseen post-rigor transition state that binds the ATP analog AppNHp. The structures reveal a high flexibility of myosin in each state and provide valuable insights into the structural transitions of myosin-V upon ADP release and binding of AppNHp, as well as the actomyosin interface. In addition, they show how myosin is able to specifically alter the structure of F-actin.
Data availability
The atomic models and cryo-EM maps are available in the PDB (Burley et al., 2018) and EMDB databases (Lawson et al., 2011), under following accession numbers: aged PHD-stabilized actomyosin-V in the strong-ADP state: 7PM5, EMD-13521 (central 1er), 7PM6, EMD-13522 (central 3er/2er), 7PM7, EMD-13523 (class 2), 7PM8, EMD-13524 (class 3), 7PM9, EMD-13525 (class 4), 7PMA, EMD-13526 (class 5), 7PMB, EMD-13527 (class 6), 7PMC, EMD-13528 (class 7) ; aged PHD-stabilized actomyosin-V in the rigor state: 7PLT, EMD-13501 (central 1er), 7PLU, EMD-13502 (central 3er/2er), 7PLV, EMD-13503 (class 1), 7PLW, EMD-13504 (class 3) and 7PLX, EMD-13505 (class 4); aged PHD-stabilized actomyosin-V in the PRT state: 7PMD, EMD-13529 (central 1er), 7PME, EMD-13530 (central 3er/2er), 7PMF, EMD-13531 (class 1), 7PMG, EMD-13532 (class 3), 7PMH, EMD-13533 (class 4), 7PMI, EMD-13535 (class 5), 7PMJ, EMD-13536 (class 6), 7PML, EMD-13538 (class 8); young JASP-stabilized actomyosin-V in the rigor state: 7PLY, EMD-13506 (central 1er), 7PLZ, EMD-13507 (central 3er/2er), 7PM0, EMD-13508 (class 1), 7PM1, EMD-13509 (class 2), 7PM2, EMD-13510 (class 4); and young JASP-stabilized F-actin: 7PM3, EMD-13511. The datasets generated during the current study are available from the corresponding author upon reasonable request.
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Cryo-EM structure of the actomyosin-V complex in the strong-ADP state (central 1er)RCSB Protein Data Bank, 7PM5.
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Cryo-EM structure of the actomyosin-V complex in the strong-ADP state (central 3er/2er)RCSB Protein Data Bank, 7PM6.
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Cryo-EM structure of the actomyosin-V complex in the rigor state (central 1er)RCSB Protein Data Bank, 7PLT.
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Cryo-EM structure of the actomyosin-V complex in the rigor state (central 3er/2er)RCSB Protein Data Bank, 7PLU.
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Cryo-EM structure of the actomyosin-V complex in the rigor state (central 1er, class 1)RCSB Protein Data Bank, 7PLV.
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Cryo-EM structure of the actomyosin-V complex in the rigor state (central 1er, class 2)RCSB Protein Data Bank, 7PLW.
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Cryo-EM structure of the actomyosin-V complex in the rigor state (central 1er, class 4)RCSB Protein Data Bank, 7PLX.
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Cryo-EM structure of young JASP-stabilized F-actin (central 3er)RCSB Protein Data Bank, 7PM3.
Article and author information
Author details
Funding
Max-Planck-Gesellschaft
- Stefan Raunser
European Commission (ERC-2019-SyG,856118)
- Stefan Raunser
Agence Nationale de la Recherche (ANR-17-CE11-0029-01)
- Anne Houdusse
National Institutes of Health (R01-DC009100)
- H Lee Sweeney
Centre National de la Recherche Scientifique
- Anne Houdusse
Studienstiftung des Deutschen Volkes
- Sabrina Pospich
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2021, Pospich 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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Further reading
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In eukaryotes, RNAs transcribed by RNA Pol II are modified at the 5′ end with a 7-methylguanosine (m7G) cap, which is recognized by the nuclear cap binding complex (CBC). The CBC plays multiple important roles in mRNA metabolism, including transcription, splicing, polyadenylation, and export. It promotes mRNA export through direct interaction with a key mRNA export factor, ALYREF, which in turn links the TRanscription and EXport (TREX) complex to the 5′ end of mRNA. However, the molecular mechanism for CBC-mediated recruitment of the mRNA export machinery is not well understood. Here, we present the first structure of the CBC in complex with an mRNA export factor, ALYREF. The cryo-EM structure of CBC-ALYREF reveals that the RRM domain of ALYREF makes direct contact with both the NCBP1 and NCBP2 subunits of the CBC. Comparing CBC-ALYREF with other cellular complexes containing CBC and/or ALYREF components provides insights into the coordinated events during mRNA transcription, splicing, and export.
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