Use of free-flow electrophoresis for the analysis of cellular uptake of picornaviruses

Electrophoresis
P KronenbergerRenate Fuchs

Abstract

Free-flow electrophoresis is a powerful tool to separate subcellular vesicles such as early and late endosomes from plasma membranes. Using this technique, the intracellular distribution of poliovirus type 2 Sabin (PV2) and its derived subviral particles was analyzed upon infection of HeLa cells. Comparison of various infection conditions showed that maximally 30% of total cell associated PV2 was found in endosomal compartments with the remainder being associated with plasma membrane fractions; 2% of viral label was recovered from the cytoplasm in form of free virions. Sucrose gradient centrifugation analysis of the viral material recovered from the respective fractions revealed that intracellular virus was exclusively in its native conformation. This is in sharp contrast to human rhinovirus serotype 2 (HRV2), which is rapidly modified to RNA-free subviral particles upon accumulation in endosomes. The data suggest that productive poliovirus uncoating can occur at the plasma membrane whereas internalized virus is most probably aborted.

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Citations

May 12, 2005·Virus Research·Steffen MuellerJeronimo Cello
Jul 12, 2007·Virology Journal·Nidia H De Jesus
Jul 14, 1998·Electrophoresis·L Krivánková, P Bocek
Oct 8, 2011·International Journal of Pharmaceutics·Mamta KapoorSiddhesh D Patil
Jul 16, 2010·Reviews in Medical Virology·Renate Fuchs, Dieter Blaas
Feb 22, 2005·The Journal of General Virology·Denise Egger, Kurt Bienz

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