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Design and self-assembly of simple coat proteins for artificial viruses

Nature Nanotechnology volume 9, pages 698–702 (2014)Cite this article

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Abstract

Viruses are among the simplest biological systems and are highly effective vehicles for the delivery of genetic material into susceptible host cells1. Artificial viruses can be used as model systems for providing insights into natural viruses and can be considered a testing ground for developing artificial life. Moreover, they are used in biomedical and biotechnological applications, such as targeted delivery of nucleic acids for gene therapy1,2 and as scaffolds in material science3,4,5. In a natural setting, survival of viruses requires that a significant fraction of the replicated genomes be completely protected by coat proteins. Complete protection of the genome is ensured by a highly cooperative supramolecular process between the coat proteins and the nucleic acids, which is based on reversible, weak and allosteric interactions only6,7,8,9. However, incorporating this type of supramolecular cooperativity into artificial viruses remains challenging10,11,12,13,14,15. Here, we report a rational design for a self-assembling minimal viral coat protein based on simple polypeptide domains. Our coat protein features precise control over the cooperativity of its self-assembly with single DNA molecules to finally form rod-shaped virus-like particles. We confirm the validity of our design principles by showing that the kinetics of self-assembly of our virus-like particles follows a previous model developed for tobacco mosaic virus9. We show that our virus-like particles protect DNA against enzymatic degradation and transfect cells with considerable efficiency, making them promising delivery vehicles.

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Figure 1: Design of the minimal viral coat protein C–Sn–B.
Figure 2: Self-assembly of VLPs: AFM and cryo-TEM images of complexes of linear dsDNA with C–Sn–B show morphologies that depend on the size n of the self-assembly block.
Figure 3: Cooperativity of the self-assembly of VLPs.
Figure 4: Formation process of VLPs.

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References

  1. Naldini, L. et al. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 272, 263–267 (1996).

    Article  CAS  Google Scholar 

  2. Mastrobattista, E., van der Aa, M. A. E. M., Hennink, W. E. & Crommelin, D. J. A. Artificial viruses: a nanotechnological approach to gene delivery. Nature Rev. Drug Discov. 5, 115–121 (2006).

    Article  Google Scholar 

  3. Comellas-Aragones, M. et al. A virus-based single-enzyme nanoreactor. Nature Nanotech. 2, 635–639 (2007).

    Article  CAS  Google Scholar 

  4. Grigoryan, G. et al. Computational design of virus-like protein assemblies on carbon nanotube surfaces. Science 332, 1071–1076 (2011).

    Article  CAS  Google Scholar 

  5. Douglas, T. & Young, M. Host–guest encapsulation of materials by assembled virus protein cages. Nature 393, 152–155 (1998).

    Article  CAS  Google Scholar 

  6. Klug, A. The tobacco mosaic virus particle: structure and assembly. Phil. Trans. R. Soc. Lond. B 354, 531–535 (1999).

    Article  CAS  Google Scholar 

  7. Caspar, D. L. & Namba, K. Switching in the self-assembly of tobacco mosaic virus. Adv. Biophys. 26, 157–185 (1990).

    Article  CAS  Google Scholar 

  8. Butler, P. J. G. Self–assembly of tobacco mosaic virus: the role of an intermediate aggregate in generating both specificity and speed. Phil. Trans. R. Soc. Lond. B 354, 537–550 (1999).

    Article  CAS  Google Scholar 

  9. Kraft, D. J., Kegel, W. K. & van der Schoot, P. A kinetic zipper model and the assembly of tobacco mosaic virus. Biophys. J. 102, 2845–2855 (2012).

    Article  CAS  Google Scholar 

  10. Percec, V., Heck, J., Johansson, G., Tomazos, D. & Ungar, G. Towards tobacco mosaic virus-like self-assembled supramolecular architectures. Macromol. Symp. 77, 237–265 (1994).

    Article  CAS  Google Scholar 

  11. Remy, J. S., Kichler, A., Mordvinov, V., Schuber, F. & Behr, J. P. Targeted gene transfer into hepatoma cells with lipopolyamine-condensed DNA particles presenting galactose ligands: a stage toward artificial viruses. Proc. Natl Acad. Sci. USA 92, 1744–1748 (1995).

    Article  CAS  Google Scholar 

  12. Aoyama, Y. et al. Artificial viruses and their application to gene delivery. Size-controlled gene coating with glycocluster nanoparticles. J. Am. Chem. Soc. 125, 3455–3457 (2003).

    Article  CAS  Google Scholar 

  13. Wagner, E. Strategies to improve DNA polyplexes for in vivo gene transfer: will ‘artificial viruses’ be the answer? Pharm. Res. 21, 8–14 (2004).

    Article  CAS  Google Scholar 

  14. Lim, Y. B., Lee, E., Yoon, Y. R., Lee, M. S. & Lee, M. Filamentous artificial virus from a self-assembled discrete nanoribbon. Angew. Chem. Int. Ed. 47, 4525–4528 (2008).

    Article  CAS  Google Scholar 

  15. Miyata, K., Nishiyama, N. & Kataoka, K. Rational design of smart supramolecular assemblies for gene delivery: chemical challenges in the creation of artificial viruses. Chem. Soc. Rev. 41, 2562–2574 (2012).

    Article  CAS  Google Scholar 

  16. Whitty, A. Cooperativity and biological complexity. Nature Chem. Biol. 4, 435–439 (2008).

    Article  CAS  Google Scholar 

  17. Namba, K., Pattanayek, R. & Stubbs, G. Visualization of protein–nucleic acid interactions in a virus. Refined structure of intact tobacco mosaic virus at 2.9 Å resolution by X-ray fiber diffraction. J. Mol. Biol. 208, 307–325 (1989).

    Article  CAS  Google Scholar 

  18. Hernandez-Garcia, A., Werten, M. W., Stuart, M. C., de Wolf, F. A. & de Vries R. Coating of single DNA molecules by genetically engineered protein diblock copolymers. Small 8, 3491–3501 (2012).

    Article  CAS  Google Scholar 

  19. Krejchi, M. T. et al. Chemical sequence control of beta-sheet assembly in macromolecular crystals of periodic polypeptides. Science 265, 1427–1432 (1994).

    Article  CAS  Google Scholar 

  20. Beun, L. H., Beaudoux, X. J., Kleijn, J. M., de Wolf, F. A. & Stuart, M. A. C. Self-assembly of silk-collagen-like triblock copolymers resembles a supramolecular living polymerization. ACS Nano 6, 133–140 (2012).

    Article  CAS  Google Scholar 

  21. Werten, M. W. T., Wisselink, W. H., Jansen-van den Bosch, T. J., de Bruin, E. C. & de Wolf, F. A. Secreted production of a custom-designed, highly hydrophilic gelatin in Pichia pastoris. Protein Eng. 14, 447–454 (2001).

    Article  CAS  Google Scholar 

  22. DeRouchey, J., Walker, G. F., Wagner, E. & Radler, J. O. Decorated rods: a ‘bottom-up’ self-assembly of monomolecular DNA complexes. J. Phys. Chem. B 110, 4548–4554 (2006).

    Article  CAS  Google Scholar 

  23. Osada, K. et al. Quantized folding of plasmid DNA condensed with block catiomer into characteristic rod structures promoting transgene efficacy. J. Am. Chem. Soc. 132, 12343–12348 (2010).

    Article  CAS  Google Scholar 

  24. Ruff, Y., Moyer, T., Newcomb, C. J., Demeler, B. & Stupp, S. I. Precision templating with DNA of a virus-like particle with peptide nanostructures. J. Am. Chem. Soc. 135, 6211–6219 (2013).

    Article  CAS  Google Scholar 

  25. Lecuyer, K. A., Behlen, L. S. & Uhlenbeck, O. C. Mutants of the bacteriophage MS2 coat protein that alter its cooperative binding to RNA. Biochemistry 34, 10600–10606 (1995).

    Article  CAS  Google Scholar 

  26. Williamson, J. R. Cooperativity in macromolecular assembly. Nature Chem. Biol. 4, 458–465 (2008).

    Article  CAS  Google Scholar 

  27. Morrone, S. R. et al. Cooperative assembly of IFI16 filaments on dsDNA provides insights into host defense strategy. Proc. Natl Acad. Sci. USA 111, E62 (2014).

    Article  CAS  Google Scholar 

  28. Rabotyagova, O. S., Cebe, P. & Kaplan, D. L. Protein-based block copolymers. Biomacromolecules 12, 269–289 (2011).

    Article  CAS  Google Scholar 

  29. Zhang, S. G. Fabrication of novel biomaterials through molecular self-assembly. Nature Biotechnol. 21, 1171–1178 (2003).

    Article  CAS  Google Scholar 

  30. King, N. P. et al. A general computational method is used to design protein building blocks that self-assemble into target architectures. Science 336, 1171–1174 (2012).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors thank A. Westphal and I. van Hees for help with the fluorescence correlation spectroscopy experiments and R. Fokkink for help with static light scattering. The authors also thank M.W.P. van de Put for help in performing part of the cryo-TEM experiments and W. Kegel for suggesting the electrophoretic light scattering experiment. A.H-G. is financially supported by the Dutch Polymer Institute (DPI), project #698 SynProt, and by the Consejo Nacional de Ciencia y Tecnología (CONACyT), México. M.E.F. is supported by the Dutch Polymer Institute (DPI, Technology area HTE, project #730). M.C.S. is supported by the European Research Council (advanced grant no. 267254). D.J.K. acknowledges financial support through a Rubicon fellowship (grant no. 680-50-1019) from the Netherlands Organization for Scientific Research (NWO).

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  1. Armando Hernandez-Garcia

    Present address: Present address: Institute for BioNanotechnology in Medicine, Northwestern University, Chicago, Illinois 60611, USA,

Authors and Affiliations

  1. Laboratory of Physical Chemistry and Colloid Science, Wageningen University, Dreijenplein 6, Wageningen, 6703 HB, The Netherlands

    Armando Hernandez-Garcia, Anne F. J. Janssen, Martien Cohen Stuart & Renko de Vries

  2. Dutch Polymer Institute, John F. Kennedylaan 2, Eindhoven, 5612 AB, The Netherlands

    Armando Hernandez-Garcia & Marco E. Favretto

  3. Soft Matter Physics, Huygens-Kamerlingh Onnes Laboratory, Leiden University, Leiden, PO Box 9504, 2300 RA, The Netherlands

    Daniela J. Kraft

  4. Department of Physics, Center for Soft Matter Research, New York University, 4 Washington Place, New York, New York 10003, USA

    Daniela J. Kraft

  5. Department of Chemical Engineering and Chemistry, Laboratory of Materials and Interface Chemistry & Soft Matter CryoTEM Research Unit, and Institute for Complex Molecular Systems, Eindhoven University of Technology, Eindhoven, PO Box 513, 5600 MB, The Netherlands

    Paul H. H. Bomans & Nico A. J. M. Sommerdijk

  6. Utrecht University, Van't Hoff Laboratory for Physical and Colloid Chemistry, Debye Institute of Nanomaterials Science, Utrecht, PO Box 80.051, 3508 TB, The Netherlands

    Dominique M. E. Thies-Weesie

  7. Department of Biochemistry, Radboud Institute of Molecular Life Sciences, Radboud University Medical Centre, Geert Grooteplein 28, Nijmegen, 6525 GA, The Netherlands

    Marco E. Favretto & Roland Brock

  8. Wageningen UR Food & Biobased Research, Bornse Weilanden 9, Wageningen, 6708 WG, The Netherlands

    Frits A. de Wolf & Marc W. T. Werten

  9. Department of Applied Physics, Theory of Polymers and Soft Matter, Eindhoven University of Technology, Eindhoven, PO Box 513, 5600 MB, The Netherlands

    Paul van der Schoot

  10. Institute for Theoretical Physics, Utrecht University, Leuvenlaan 4, Utrecht, 3584 CE, The Netherlands

    Paul van der Schoot

  11. Department of Biomedical Engineering, University Medical Center Groningen, University of Groningen, Groningen, PO Box 196, 9700 AD, The Netherlands

    Renko de Vries

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Contributions

F.d.W., M.C.S., P.v.d.S. and R.d.V. conceived the initial idea. R.d.V. and A.H-G. conceived and designed the molecules and experiments. A.H-G. produced the proteins, except for C–S2–B, which was produced by A.F.J.J. M.W. contributed to the production of proteins and to writing the technical sections on protein production. Except for the transfection experiments, analytical ultracentrifugation and cryo-TEM imaging, A.H-G. performed all experiments. A.H-G. and R.d.V. interpreted and analysed all experimental data except for the transfection experiments. P.v.d.S. and D.J.K. performed the theoretical analysis of the VLP assembly kinetics. R.B. designed the transfection experiments. M.F. performed and analysed the transfection experiments. D.M.E.T-W. designed, performed and analysed the analytical ultracentrifugation experiments. N.A.J.M.S. designed the cryo-TEM imaging experiments. P.H.H.B. performed the cryo-TEM imaging. A.H-G., D.J.K., P.v.d.S., M.C.S. and R.d.V. wrote the paper. All authors commented on the manuscript.

Corresponding authors

Correspondence to Armando Hernandez-Garcia or Renko de Vries.

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The authors declare no competing financial interests.

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Hernandez-Garcia, A., Kraft, D., Janssen, A. et al. Design and self-assembly of simple coat proteins for artificial viruses. Nature Nanotech 9, 698–702 (2014). https://doi.org/10.1038/nnano.2014.169

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