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Investigating Liquid-Liquid Phase Separation in Virus-Generated Inclusion Bodies Using Fluorescence Recovery After Photobleaching of Fluorescently Labeled Host Proteins

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Measles and Related Morbilliviruses

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2808))

Abstract

Many negative-sense single-stranded RNA viruses within the order Mononegavirales harm humans. A common feature shared among cells infected by these viruses is the formation of subcellular membraneless structures called biomolecular condensates, also known as inclusion bodies (IBs), that form through a process called liquid-liquid phase separation (LLPS). Like many other membraneless organelles, viral IBs enrich a specific subset of viral and host proteins involved in the formation of viral particles. Elucidation of the properties and regulation of these IBs as they mature throughout the viral replication process are important for our understanding of viral replication, which may also lead to the development of alternative antiviral treatments. The protocol outlined in this chapter aims to characterize the intrinsic properties of LLPS within the measles virus (MeV, a member of Mononegavirales) IBs by using an imaging approach that fluorescently tags an IB-associated host protein. This method uses common laboratory techniques and is generalizable to any host factors as well as other viral systems.

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Authors and Affiliations

  1. The Department of Molecular, Cell, and Developmental Biology, University of California Santa Barbara, Santa Barbara, CA, USA

    Matthew B. T. Asuelime-Smith

  2. Molecular, Cellular, and Developmental Biology, University of California Santa Barbara, Santa Barbara, CA, USA

    Dzwokai Z. Ma

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  1. Matthew B. T. Asuelime-Smith
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  2. Dzwokai Z. Ma
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Correspondence to Dzwokai Z. Ma .

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Editors and Affiliations

  1. Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, Santa Barbara, CA, USA

    Dzwokai Z. Ma

  2. Department of Molecular Medicine, Mayo Clinic, Rochester, MN, USA

    Christian K. Pfaller

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Asuelime-Smith, M.B.T., Ma, D.Z. (2024). Investigating Liquid-Liquid Phase Separation in Virus-Generated Inclusion Bodies Using Fluorescence Recovery After Photobleaching of Fluorescently Labeled Host Proteins. In: Ma, D.Z., Pfaller, C.K. (eds) Measles and Related Morbilliviruses. Methods in Molecular Biology, vol 2808. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3870-5_10

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  • DOI: https://doi.org/10.1007/978-1-0716-3870-5_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3869-9

  • Online ISBN: 978-1-0716-3870-5

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