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Cryo-EM Techniques to Resolve the Structure of HSV-1 Capsid-Associated Components

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Herpes Simplex Virus

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1144))

Abstract

Electron cryo-microscopy has become a routine technique to determine the structure of biochemically purified herpes simplex virus capsid particles. This chapter describes the procedures of specimen preparation by cryopreservation; low dose and low temperature imaging in an electron cryo-microscope; and data processing for reconstruction. This methodology has yielded subnanometer resolution structures of the icosahedral capsid shell where α-helices and β-sheets of individual subunits can be recognized. A relaxation of the symmetry in the reconstruction steps allows us to resolve the DNA packaging protein located at one of the 12 vertices in the capsid.

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Acknowledgements

This research was supported by grants from Robert Welch Foundation (Q1242) and National Institutes of Health (P41GM103832 and R56AI075208 to W.C.; T15LM007093 through the Gulf Coast Consortia and T32GM007330 through the MSTP to R.H.R.).

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Authors and Affiliations

  1. Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, 77030, USA

    Ryan H. Rochat & Wah Chiu

  2. Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX, 77030, USA

    Corey W. Hecksel & Wah Chiu

Authors
  1. Ryan H. Rochat
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  2. Corey W. Hecksel
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  3. Wah Chiu
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Corresponding author

Correspondence to Wah Chiu .

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Editors and Affiliations

  1. Centre for Virus Research, Westmead Millennium Institute, Westmead, New South Wales, Australia

    Russell J. Diefenbach

  2. Institute of Virology, University of Zürich, Zürich, Switzerland

    Cornel Fraefel

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Rochat, R.H., Hecksel, C.W., Chiu, W. (2014). Cryo-EM Techniques to Resolve the Structure of HSV-1 Capsid-Associated Components. In: Diefenbach, R., Fraefel, C. (eds) Herpes Simplex Virus. Methods in Molecular Biology, vol 1144. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0428-0_18

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  • DOI: https://doi.org/10.1007/978-1-4939-0428-0_18

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0427-3

  • Online ISBN: 978-1-4939-0428-0

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