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

Nano Letters
Margherita MarchettiW H Roos

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.

References

Sep 12, 2002·Clinical Pharmacokinetics·Cathryn MahTerence R Flotte
May 9, 2003·Nature·Timothy J Richmond, Curt A Davey
Jul 13, 2004·Virology·Greg L CasiniDavid T Wu
Aug 13, 2004·Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences·Theo Odijk
Aug 11, 2005·Physical Review. E, Statistical, Nonlinear, and Soft Matter Physics·Yuri O Popov, Alexei V Tkachenko
Jan 3, 2006·Biophysical Journal·Roya ZandiHoward Reiss
Aug 7, 2007·Physical Review. E, Statistical, Nonlinear, and Soft Matter Physics·Tao Hu, B I Shklovskii
Nov 13, 2007·Nature Methods·Maarten C NoomGijs J L Wuite
May 20, 2008·Biophysical Journal·Anton S Petrov, Stephen C Harvey
Jul 23, 2008·Physical Review. E, Statistical, Nonlinear, and Soft Matter Physics·Theo Odijk
Dec 31, 2008·Physical Review Letters·Nam-Kyung LeeSeok-Cheol Hong
Mar 18, 2009·Methods in Enzymology·Sarah Katen, Adam Zlotnick
May 28, 2009·Proceedings of the National Academy of Sciences of the United States of America·Amélie Leforestier, Françoise Livolant
Sep 9, 2009·Advances in Colloid and Interface Science·Wojciech PlazinskiAnita Plazinska
Jun 26, 2010·Physics of Life Reviews·Kathy R ChaurasiyaMark C Williams
Jul 21, 2010·Biophysical Journal·Aleksandr Kivenson, Michael F Hagan
Jan 25, 2011·Nature Chemistry·Charlotte UetrechtAlbert J R Heck
Jun 28, 2012·Biophysical Journal·Daniela J KraftPaul van der Schoot
Aug 7, 2012·Small·Armando Hernandez-GarciaRenko de Vries
Jun 26, 2013·ELife·Jason D PerlmutterMichael F Hagan
Jan 22, 2014·Chemical Reviews·Iddo HellerGijs J L Wuite
Jan 29, 2014·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Iddo HellerErwin J G Peterman
Aug 26, 2014·Nature Nanotechnology·Armando Hernandez-GarciaRenko de Vries
Oct 8, 2014·Proceedings of the National Academy of Sciences of the United States of America·Andrea CandelliErwin J G Peterman
Nov 25, 2014·Nature Methods·Gerrit SittersGijs J L Wuite
Dec 15, 2015·Accounts of Chemical Research·Rees F GarmannWilliam M Gelbart
Dec 19, 2015·International Journal of Pharmaceutical Investigation·Ningning Yang
Apr 18, 2016·Current Opinion in Virology·Peter C DoerschukJohn E Johnson
Apr 27, 2016·The Journal of Physical Chemistry. B·Melle Tijmen Jozef Johannes Maria PunterPaul van der Schoot
Mar 23, 2017·Journal of the American Chemical Society·Hande E CingilJoris Sprakel
Sep 25, 2017·Methods in Molecular Biology·Douwe Kamsma, Gijs J L Wuite
Jan 10, 2018·Bioengineering & Translational Medicine·Marcus J RohovieJames R Swartz
Feb 21, 2018·Journal of Physics. Condensed Matter : an Institute of Physics Journal·Siyu LiRoya Zandi
Aug 8, 2018·Nature Communications·Maelenn ChevreuilGuillaume Tresset
Aug 21, 2018·Physical Biology·Jamoliddin RazzokovPaul van der Schoot

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Citations

Jan 22, 2020·Wiley Interdisciplinary Reviews. Nanomedicine and Nanobiotechnology·Pedro BuzónWouter H Roos
Oct 22, 2019·Wiley Interdisciplinary Reviews. Nanomedicine and Nanobiotechnology·Christina Wege, Claudia Koch
Nov 26, 2020·Essays in Biochemistry·Gurleen Kaur, Lisanne M Spenkelink
Mar 18, 2021·Nano Letters·Carlos Calcines-CruzArmando Hernandez-Garcia
Mar 18, 2021·Nature Reviews Physics·Robijn F BruinsmaWouter H Roos
May 1, 2021·Life·Dinh To Le, Kristian M Müller
Jun 3, 2021·Nanomaterials·Armando Hernandez-Garcia
Jun 16, 2021·Journal of Colloid and Interface Science·Rita Ungai-SalánkiBálint Szabó
Apr 3, 2020·Journal of the American Chemical Society·Roi AsorUri Raviv
Nov 4, 2021·Science Advances·Pedro BuzónWouter H Roos

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Methods Mentioned

BETA
electron
atomic force microscopy
AFM
electron microscopy
fluorescence microscopy
optical tweezers
optical
fluoresce microscopy
electrophoretic mobility shift assay
fluorescence

Software Mentioned

LABVIEW
NanoScope Analysis
MATLAB
ImageJ

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