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Optical Tweezers to Study Viruses

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Structure and Physics of Viruses

Part of the book series: Subcellular Biochemistry ((SCBI,volume 68))

Abstract

A virus is a complex molecular machine that propagates by channeling its genetic information from cell to cell. Unlike macroscopic engines, it operates in a nanoscopic world under continuous thermal agitation. Viruses have developed efficient passive and active strategies to pack and release nucleic acids. Some aspects of the dynamic behavior of viruses and their substrates can be studied using structural and biochemical techniques. Recently, physical techniques have been applied to dynamic studies of viruses in which their intrinsic mechanical activity can be measured directly. Optical tweezers are a technology that can be used to measure the force, torque and strain produced by molecular motors, as a function of time and at the single-molecule level. Thanks to this technique, some bacteriophages are now known to be powerful nanomachines; they exert force in the piconewton range and their motors work in a highly coordinated fashion for packaging the viral nucleic acid genome. Nucleic acids, whose elasticity and condensation behavior are inherently coupled to the viral packaging mechanisms, are also amenable to examination with optical tweezers. In this chapter, we provide a comprehensive analysis of this laser-based tool, its combination with imaging methods and its application to the study of viruses and viral molecules.

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Notes

  1. 1.

    Quantum dots are luminescent semiconductor nanocrystals which are used as a new class of fluorescent probes. In comparison with organic dyes and fluorescent proteins, they are brighter and exhibit both greater photostability and longer lifetimes. They can also be tuned to specific wavelengths with narrower bandwidths than fluorescent molecules.

  2. 2.

    Confocal microscopy is a high-resolution imaging technique based on a point-by-point illumination of the sample. The light source is often a focused laser beam. Lasers of several wavelengths and filters can be used to combine this technique with fluorescent imaging.

  3. 3.

    Also especially recommended for further reading are references [1, 7, 13, 24, 25] listed above.

Abbreviations

3D:

Three dimensional

AFM:

Atomic force microscopy

bp:

Base pair

Dig:

Digoxigenin

ds:

Double-stranded

GC:

Guanine-cytosine

IR:

Infrared

NA:

Numerical aperture

α-Dig:

Anti-digoxigenin

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Acknowledgements

It is a pleasure to acknowledge J.R. Moffitt and J.L. Carrascosa for technical insights into different aspects of the chapter, R. Bocanegra, L. Quintana for careful reading of the manuscript, C. Mark and S. Hormeño for editorial and illustration assistance, respectively, and M. de la Guía for graphic design of Fig. 9.8. This work was funded by the Spanish Ministry of Science and Innovation under the “Ramon y Cajal” program (Grant No. RYC-2007-01765).

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Authors and Affiliations

  1. Instituto Madrileño de Estudios Avanzados en Nanociencia (IMDEA Nanociencia), c/Faraday 9, Campus de Cantoblanco, 28049, Madrid, Spain

    J. Ricardo Arias-Gonzalez

  2. Department of Macromolecular Structure, Centro Nacional de Biotecnología (CSIC), c/Darwin 3, Campus de Cantoblanco, 28049, Madrid, Spain

    J. Ricardo Arias-Gonzalez

  3. CNB-CSIC-IMDEA Nanociencia associated unit “Unidad de Nanobiotecnología”, Campus de Cantoblanco, 28049, Madrid, Spain

    J. Ricardo Arias-Gonzalez

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  1. J. Ricardo Arias-Gonzalez
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Correspondence to J. Ricardo Arias-Gonzalez .

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  1. "Severo Ochoa" (CSIC_UAM), And Dept. of Molecular Biology, Centro de Biología Molecular, Cantoblanco, Madrid, 28049, Madrid, Spain

    Mauricio G. Mateu

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Arias-Gonzalez, J.R. (2013). Optical Tweezers to Study Viruses. In: Mateu, M. (eds) Structure and Physics of Viruses. Subcellular Biochemistry, vol 68. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-6552-8_9

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