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Rapid, Seamless Generation of Recombinant Poxviruses using Host Range and Visual Selection

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Published on: May 24, 2020

Poxvirus membrane biogenesis: rupture not disruption.

Jacomine Krijnse Locker¹, Petr Chlanda, Timo Sachsenheimer

¹Electron Microscopy Core Facility & Department of Infectious Diseases, Heidelberg University, Im Neuenheimer Feld 267, 69120 Heidelberg, Germany. jacomine.krijnse@bioquant.uni-heidelberg.de

Cellular Microbiology

|November 22, 2012

Summary

This study reveals vaccinia virus (VACV) creates its own membrane through a novel rupture process, not host budding. Lipid analysis suggests an endoplasmic reticulum origin for the viral membrane.

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Methodology for the Efficient Generation of Fluorescently Tagged Vaccinia Virus Proteins

Methodology for the Efficient Generation of Fluorescently Tagged Vaccinia Virus Proteins

Published on: January 17, 2014

Area of Science:

Virology
Cell Biology
Biochemistry

Background:

Enveloped viruses typically acquire membranes from host cells via budding.
Previous transmission electron microscopy (TEM) suggested vaccinia virus (VACV) might form its membrane de novo.

Purpose of the Study:

To investigate the biogenesis and origin of the VACV membrane.
To elucidate the unconventional mechanism of viral membrane formation.

Main Methods:

Electron tomography (ET) and cryo-electron microscopy (cryo-EM) were used to visualize VACV.
Mass spectrometry (MS) was employed for lipid analysis of purified VACV.

Main Results:

Data support a model of unconventional membrane biogenesis involving membrane rupture and open membrane intermediates.
Lipid analysis indicates an endoplasmic reticulum (ER) origin, with lower cholesterol than host cells.
Enrichment of phosphatidic acid, diacylglycerol, and phosphatidylinositol was detected in the virion.

Conclusions:

VACV utilizes a unique membrane biogenesis pathway involving rupture, distinct from typical viral budding.
The findings contribute to understanding viral membrane origins and potential implications for other pathogens.