Real-Time Assembly of Viruslike Nucleocapsids Elucidated at the Single-Particle Level

Margherita Marchetti, Douwe Kamsma, Ernesto Cazares Vargas, Armando Hernandez García, Paul Van Der Schoot, Renko De Vries, Gijs J.L. Wuite, Wouter H. Roos

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

While the structure of a multitude of viral particles has been resolved to atomistic detail, their assembly pathways remain largely elusive. Key unresolved issues are particle nucleation, particle growth, and the mode of genome compaction. These issues are difficult to address in bulk approaches and are effectively only accessible by the real-time tracking of assembly dynamics of individual particles. This we do here by studying the assembly into rod-shaped viruslike particles (VLPs) of artificial capsid polypeptides. Using fluorescence optical tweezers, we establish that small oligomers perform one-dimensional diffusion along the DNA. Larger oligomers are immobile and nucleate VLP growth. A multiplexed acoustic force spectroscopy approach reveals that DNA is compacted in regular steps, suggesting packaging via helical wrapping into a nucleocapsid. By reporting how real-time assembly tracking elucidates viral nucleation and growth principles, our work opens the door to a fundamental understanding of the complex assembly pathways of both VLPs and naturally evolved viruses.

Original languageEnglish
Pages (from-to)5746-5753
Number of pages8
JournalNano Letters
Volume19
Issue number8
Early online date1 Aug 2019
DOIs
Publication statusPublished - 14 Aug 2019

Fingerprint

Nucleocapsid
assembly
Oligomers
Packaging
DNA
Nucleation
oligomers
Optical tweezers
Polypeptides
deoxyribonucleic acid
nucleation
Viruses
genome
Compaction
polypeptides
Genes
viruses
Acoustics
Fluorescence
Spectroscopy

Keywords

  • acoustic force spectroscopy
  • artificial virus
  • biophysics
  • optical tweezers
  • physical virology
  • Self-assembly

Cite this

Marchetti, M., Kamsma, D., Cazares Vargas, E., Hernandez García, A., Van Der Schoot, P., De Vries, R., ... Roos, W. H. (2019). Real-Time Assembly of Viruslike Nucleocapsids Elucidated at the Single-Particle Level. Nano Letters, 19(8), 5746-5753. https://doi.org/10.1021/acs.nanolett.9b02376
Marchetti, Margherita ; Kamsma, Douwe ; Cazares Vargas, Ernesto ; Hernandez García, Armando ; Van Der Schoot, Paul ; De Vries, Renko ; Wuite, Gijs J.L. ; Roos, Wouter H. / Real-Time Assembly of Viruslike Nucleocapsids Elucidated at the Single-Particle Level. In: Nano Letters. 2019 ; Vol. 19, No. 8. pp. 5746-5753.
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Marchetti, M, Kamsma, D, Cazares Vargas, E, Hernandez García, A, Van Der Schoot, P, De Vries, R, Wuite, GJL & Roos, WH 2019, 'Real-Time Assembly of Viruslike Nucleocapsids Elucidated at the Single-Particle Level' Nano Letters, vol. 19, no. 8, pp. 5746-5753. https://doi.org/10.1021/acs.nanolett.9b02376

Real-Time Assembly of Viruslike Nucleocapsids Elucidated at the Single-Particle Level. / Marchetti, Margherita; Kamsma, Douwe; Cazares Vargas, Ernesto; Hernandez García, Armando; Van Der Schoot, Paul; De Vries, Renko; Wuite, Gijs J.L.; Roos, Wouter H.

In: Nano Letters, Vol. 19, No. 8, 14.08.2019, p. 5746-5753.

Research output: Contribution to JournalArticleAcademicpeer-review

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Marchetti M, Kamsma D, Cazares Vargas E, Hernandez García A, Van Der Schoot P, De Vries R et al. Real-Time Assembly of Viruslike Nucleocapsids Elucidated at the Single-Particle Level. Nano Letters. 2019 Aug 14;19(8):5746-5753. https://doi.org/10.1021/acs.nanolett.9b02376