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Stimulus-responsive assembly of nonviral nucleocapsids

Author

Listed:
  • Mao Hori

    (ETH Zürich
    Tokyo Medical and Dental University)

  • Angela Steinauer

    (ETH Zürich
    SB ISIC LIBN)

  • Stephan Tetter

    (ETH Zürich
    Francis Crick Avenue)

  • Jamiro Hälg

    (ETH Zürich)

  • Eva-Maria Manz

    (ETH Zürich)

  • Donald Hilvert

    (ETH Zürich)

Abstract

Controlled assembly of a protein shell around a viral genome is a key step in the life cycle of many viruses. Here we report a strategy for regulating the co-assembly of nonviral proteins and nucleic acids into highly ordered nucleocapsids in vitro. By fusing maltose binding protein to the subunits of NC-4, an engineered protein cage that encapsulates its own encoding mRNA, we successfully blocked spontaneous capsid assembly, allowing isolation of the individual monomers in soluble form. To initiate RNA-templated nucleocapsid formation, the steric block can be simply removed by selective proteolysis. Analyses by transmission and cryo-electron microscopy confirmed that the resulting assemblies are structurally identical to their RNA-containing counterparts produced in vivo. Enzymatically triggered cage formation broadens the range of RNA molecules that can be encapsulated by NC-4, provides unique opportunities to study the co-assembly of capsid and cargo, and could be useful for studying other nonviral and viral assemblies.

Suggested Citation

  • Mao Hori & Angela Steinauer & Stephan Tetter & Jamiro Hälg & Eva-Maria Manz & Donald Hilvert, 2024. "Stimulus-responsive assembly of nonviral nucleocapsids," Nature Communications, Nature, vol. 15(1), pages 1-10, December.
    • Handle: RePEc:nat:natcom:v:15:y:2024:i:1:d:10.1038_s41467-024-47808-1
    DOI: 10.1038/s41467-024-47808-1
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    References listed on IDEAS

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    1. Yang Hsia & Jacob B. Bale & Shane Gonen & Dan Shi & William Sheffler & Kimberly K. Fong & Una Nattermann & Chunfu Xu & Po-Ssu Huang & Rashmi Ravichandran & Sue Yi & Trisha N. Davis & Tamir Gonen & Nei, 2016. "Design of a hyperstable 60-subunit protein icosahedron," Nature, Nature, vol. 535(7610), pages 136-139, July.
    2. Eita Sasaki & Daniel Böhringer & Michiel van de Waterbeemd & Marc Leibundgut & Reinhard Zschoche & Albert J. R. Heck & Nenad Ban & Donald Hilvert, 2017. "Structure and assembly of scalable porous protein cages," Nature Communications, Nature, vol. 8(1), pages 1-10, April.
    3. Eyal Golub & Rohit H. Subramanian & Julian Esselborn & Robert G. Alberstein & Jake B. Bailey & Jerika A. Chiong & Xiaodong Yan & Timothy Booth & Timothy S. Baker & F. Akif Tezcan, 2020. "Constructing protein polyhedra via orthogonal chemical interactions," Nature, Nature, vol. 578(7793), pages 172-176, February.
    4. Robert P. King, 2012. "The Science of Design," American Journal of Agricultural Economics, Agricultural and Applied Economics Association, vol. 94(2), pages 275-284.
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