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

Publisher: Humana Press Country of Publication: United States NLM ID: 9214969 Publication Model: Print Cited Medium: Internet ISSN: 1940-6029 (Electronic) Linking ISSN: 10643745 NLM ISO Abbreviation: Methods Mol Biol Subsets: MEDLINE

Publication: Totowa, NJ : Humana Press
Original Publication: Clifton, N.J. : Humana Press,

Inclusion Bodies, Viral*/metabolism ; Fluorescence Recovery After Photobleaching*/methods ; Measles virus*/physiology ; Measles virus*/metabolism; Humans ; Virus Replication ; Inclusion Bodies/metabolism ; Animals ; Host-Pathogen Interactions ; Phase Separation

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

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Keywords: FRAP; Fluorescence recovery after photobleaching; Host factors; Innate immune response; LLPS; Liquid-liquid phase separation; Measles virus; Mononegavirales; Paramyxovirus; Viral inclusion bodies

Date Created: 20240514 Date Completed: 20240514 Latest Revision: 20240514

20240515

10.1007/978-1-0716-3870-5_10

38743367