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Imaging the alphavirus exit pathway.

Maria Guadalupe Martinez¹, Erik-Lee Snapp², Geoffrey S Perumal³

¹Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York, USA.

Journal of Virology

|April 4, 2014

Summary

This summary is machine-generated.

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Alphavirus infection reorganizes the host cell plasma membrane, creating specialized sites for virus assembly and budding. This process involves viral proteins excluding host proteins and utilizing cellular extensions for particle release and transfer.

Area of Science:

Virology
Cell Biology
Structural Biology

Background:

Alphaviruses are enveloped RNA viruses that bud from the host cell plasma membrane (PM).
The precise mechanisms and dynamics of alphavirus assembly and budding remain poorly understood.
Alphaviruses are significant human pathogens necessitating the development of vaccines and antiviral therapies.

Purpose of the Study:

To characterize the dynamics of alphavirus assembly and budding at the host cell PM.
To investigate the role of viral envelope proteins in organizing budding sites.
To elucidate the relationship between viral structure, host cell reorganization, and virus release.

Main Methods:

Generation of Sindbis viruses (SINVs) with fluorescent protein-tagged E2 envelope proteins.

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Alphavirus Transducing System: Tools for Visualizing Infection in Mosquito Vectors

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Live Cell Imaging of Alphaherpes Virus Anterograde Transport and Spread

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Live-cell imaging using fluorescence microscopy to track viral and host proteins.

Correlative light and electron microscopy (CLEM) to link molecular events with ultrastructure.

Analysis of host cell plasma membrane protein exclusion and cytoskeletal involvement.

Main Results:

Alphavirus infection induced localized patches and filopodium-like extensions on the PM enriched with viral E2 protein.
These viral protein-rich sites excluded host PM marker proteins, indicating specialized membrane domains.
Exclusion of host proteins was dependent on the interaction between viral E2 and capsid proteins.
Two types of extensions were observed: short, PM-excluding extensions and long, PM-containing extensions capable of virus transfer.
Tubulin-positive extensions were crucial for virus particle transfer and were reduced in nonbudding mutants.

Conclusions:

Alphavirus infection triggers significant reorganization of the host cell plasma membrane and cytoskeleton.
Virus assembly and budding occur at specialized, virus-induced membrane domains that exclude host proteins.
The interaction between viral capsid and envelope proteins is critical for forming these specialized budding sites.
Cytoskeletal elements, particularly tubulin, play a role in virus particle transfer via cellular extensions.