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A Quantitative Fusion Assay to Study Measles Virus Entry

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Measles and Related Morbilliviruses

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

Abstract

We have adopted a real-time assay based on a dual-split reporter to assess cell-cell fusion mediated by the measles virus (MeV) membrane fusion machinery. This reporter system is comprised of two expression vectors, each encoding a segment of Renilla luciferase fused to a segment of GFP. To regain function, the two segments need to associate, which is dependent on cell-cell fusion between effector cells expressing the MeV fusion machinery and target cells expressing the corresponding MeV receptor. By measuring reconstituted luciferase activity, we can follow the kinetics of cell-cell fusion and quantify the extent of fusion. This assay lends itself to the study of the MeV fusion machinery comprised of the attachment and fusion glycoproteins, the matrix protein, and the MeV receptors. Moreover, entry inhibitors targeting attachment or fusion can be readily screened using this assay. Finally, this assay can be easily adopted to study the entry of other members of the Paramyxoviridae, as we have demonstrated for the henipaviruses.

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References

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Authors and Affiliations

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

    Chanakha K. Navaratnarajah

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  1. Chanakha K. Navaratnarajah
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Correspondence to Chanakha K. Navaratnarajah .

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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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Navaratnarajah, C.K. (2024). A Quantitative Fusion Assay to Study Measles Virus Entry. 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_1

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  • DOI: https://doi.org/10.1007/978-1-0716-3870-5_1

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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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