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J Virol, February 1998, p. 1354-1364, Vol. 72, No. 2
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Major and Minor Receptor Group Human Rhinoviruses Penetrate from Endosomes by Different Mechanisms

Daniela Schober,1 Peter Kronenberger,2 Elisabeth Prchla,1 Dieter Blaas,3 and Renate Fuchs1,*

Department of General and Experimental Pathology, University of Vienna, A-1090 Vienna,1 and Institute of Biochemistry, University of Vienna, A-1030 Vienna,3 Austria, and Department of Microbiology and Hygiene, Vrije Universiteit Brussel, B-1090 Brussels, Belgium2

Received 14 February 1997/Accepted 25 October 1997

Intercellular adhesion molecule 1 and the low-density lipoprotein receptor are used for cell entry by major and minor receptor group human rhinoviruses (HRVs), respectively. Whereas minor-group viruses, exemplified by HRV2, transfer their genomic RNA to the cytoplasm through a pore in the endosomal membrane (E. Prchla, C. Plank, E. Wagner, D. Blaas, and R. Fuchs, J. Cell Biol. 131:111-123, 1995), the mechanism of in vivo uncoating of major-group HRVs has not been elucidated so far. Using free-flow electrophoresis, we performed a comparative analysis of cell entry by HRV2 and the major group rhinovirus HRV14. Here we demonstrate that this technique allows the separation of free viral particles from those associated with early endosomes, late endosomes, and plasma membranes. Upon free-flow electrophoretic separation of microsomes, HRV14 was recovered from endosomes under conditions which prevent uncoating, whereas the proportion of free viral particles increased with time under conditions which promote uncoating. The remaining virus eluted within numerous fractions corresponding to membraneous material, with no clear endosomal peaks being discernible. This suggests that uncoating of HRV14 results in lysis of the endosomal membrane and release of subviral 135S and 80S particles into the cytoplasm.

* Corresponding author. Mailing address: Department of General and Experimental Pathology, University of Vienna, Währinger Gürtel 18-20, A-1090 Vienna, Austria. Phone: (43 1) 40 400 5127. Fax: (43 1) 40 400 5130. E-mail: renate.fuchs{at}akh-wien.ac.at




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