This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Berka, U.
Right arrow Articles by Fuchs, R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Berka, U.
Right arrow Articles by Fuchs, R.

 Previous Article  |  Next Article 

Journal of Virology, April 2009, p. 3778-3787, Vol. 83, No. 8
0022-538X/09/$08.00+0     doi:10.1128/JVI.01739-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Human Rhinovirus Type 2 Uncoating at the Plasma Membrane Is Not Affected by a pH Gradient but Is Affected by the Membrane Potential{triangledown}

Ursula Berka,1 Abdul Khan,2 Dieter Blaas,2 and Renate Fuchs1*

Department of Pathophysiology, and Max F. Perutz Laboratories, University Departments at the Vienna Biocenter,1 Department of Medical Biochemistry, Medical University of Vienna, Vienna, Austria2

Received 17 August 2008/ Accepted 22 January 2009

The minor receptor group human rhinovirus type 2 enters host cells by endocytosis via members of the low-density-lipoprotein receptor family. In late endosomes, it undergoes a conformational change solely induced by a pH of ≤5.6, resulting in RNA transfer across the endosomal membrane into the cytoplasm. To determine potential driving forces of this process, we investigated whether RNA penetration might depend on the pH gradient and/or the membrane potential between the acidic endosome lumen and the neutral cytoplasm. Since these parameters are difficult to assess in endosomes, we took advantage of the possibility of inducing structural changes, RNA release, and consequently infection from the plasma membrane. To manipulate the pH gradient, cell-bound virus was exposed to membrane-permeant or -impermeant acidic buffers at 4°C, and this was followed by a shift to 34°C in medium containing bafilomycin to prevent RNA release from endosomes. To manipulate the plasma membrane potential, similar experiments were carried out, but these included K+ diffusion potentials in the presence of the K+ ionophore valinomycin. We demonstrated that infection does not depend on a pH gradient but is enhanced by plasma membrane hyperpolarization compared to plasma membrane depolarization.

* Corresponding author. Mailing address: Department of Pathophysiology, Medical 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}meduniwien.ac.at

{triangledown} Published ahead of print on 4 February 2009.

Journal of Virology, April 2009, p. 3778-3787, Vol. 83, No. 8
0022-538X/09/$08.00+0     doi:10.1128/JVI.01739-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»