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 HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Imelli, N.
Right arrow Articles by Greber, U. F.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Imelli, N.
Right arrow Articles by Greber, U. F.

 Previous Article  |  Next Article 

Journal of Virology, March 2004, p. 3089-3098, Vol. 78, No. 6
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.6.3089-3098.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Cholesterol Is Required for Endocytosis and Endosomal Escape of Adenovirus Type 2

Nicola Imelli,1 Oliver Meier,1 Karin Boucke,1 Silvio Hemmi,2 and Urs F. Greber1*

Zoologisches Institut,1 Institut für Molekularbiologie, Universität Zürich, CH-8057 Zürich, Switzerland2

Received 19 September 2003/ Accepted 20 November 2003

The species C adenovirus type 2 (Ad2) and Ad5 bind the coxsackievirus B Ad receptor and {alpha}v integrin coreceptors and enter epithelial cells by clathrin-mediated endocytosis. This pathway is rapid and efficient. It leads to cell activation and the cholesterol-dependent formation of macropinosomes. Macropinosomes are triggered to release their contents when incoming Ad2 escapes from endosomes. Here, we show that cholesterol extraction of epithelial cells by methyl-ß-cyclodextrin (mßCD) treatment reduced Ad5-mediated luciferase expression ~4-fold. The addition of cholesterol to normal cells increased gene expression in a dose-dependent manner up to threefold, but it did not restore gene expression in mßCD-treated cells. mßCD had no effect in the presence of excess cholesterol, indicating that the inhibition of gene expression was due specifically to cholesterol depletion. Cholesterol depletion inhibited rapid Ad2 endocytosis, endosomal escape, and nuclear targeting, consistent with the notion that clathrin-dependent endocytosis of Ad2 is cholesterol dependent. In cholesterol-reduced cells, Ad2 internalized at a low rate, suggestive of an alternative, clathrin-independent, low-capacity entry pathway. While exogenous cholesterol completely restored rapid Ad2 endocytosis, macropinocytosis, and macropinosome disruption, it did not, surprisingly, restore viral escape from endosomes. Our results indicate that macropinosome disruption and endosomal escape of Ad2 are independent events in cells depleted of and then refilled with cholesterol, suggesting that viral escape from endosomes requires lipid-controlled membrane homeostasis, trafficking, or signaling.

* Corresponding author. Mailing address: Zoologisches Institut der Universität Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland. Phone: 41 1 635 4841. Fax: 41 1 635 6822. E-mail: ufgreber{at}zool.unizh.ch.

Journal of Virology, March 2004, p. 3089-3098, Vol. 78, No. 6
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.6.3089-3098.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Cianciola, N. L., Carlin, C. R. (2009). Adenovirus RID-{alpha} activates an autonomous cholesterol regulatory mechanism that rescues defects linked to Niemann-Pick disease type C. JCB 187: 537-552 [Abstract] [Full Text]  
  • Martin-Acebes, M. A., Gonzalez-Magaldi, M., Vazquez-Calvo, A., Armas-Portela, R., Sobrino, F. (2009). Internalization of Swine Vesicular Disease Virus into Cultured Cells: a Comparative Study with Foot-and-Mouth Disease Virus. J. Virol. 83: 4216-4226 [Abstract] [Full Text]  
  • Simon, M., Johansson, C., Mirazimi, A. (2009). Crimean-Congo hemorrhagic fever virus entry and replication is clathrin-, pH- and cholesterol-dependent. J. Gen. Virol. 90: 210-215 [Abstract] [Full Text]  
  • Funk, A., Mhamdi, M., Hohenberg, H., Heeren, J., Reimer, R., Lambert, C., Prange, R., Sirma, H. (2008). Duck Hepatitis B Virus Requires Cholesterol for Endosomal Escape during Virus Entry. J. Virol. 82: 10532-10542 [Abstract] [Full Text]  
  • Vassilieva, E. V., Gerner-Smidt, K., Ivanov, A. I., Nusrat, A. (2008). Lipid rafts mediate internalization of {beta}1-integrin in migrating intestinal epithelial cells. Am. J. Physiol. Gastrointest. Liver Physiol. 295: G965-G976 [Abstract] [Full Text]  
  • Inoue, Y., Tanaka, N., Tanaka, Y., Inoue, S., Morita, K., Zhuang, M., Hattori, T., Sugamura, K. (2007). Clathrin-Dependent Entry of Severe Acute Respiratory Syndrome Coronavirus into Target Cells Expressing ACE2 with the Cytoplasmic Tail Deleted. J. Virol. 81: 8722-8729 [Abstract] [Full Text]  
  • Raghu, H., Sharma-Walia, N., Veettil, M. V., Sadagopan, S., Caballero, A., Sivakumar, R., Varga, L., Bottero, V., Chandran, B. (2007). Lipid Rafts of Primary Endothelial Cells Are Essential for Kaposi's Sarcoma-Associated Herpesvirus/Human Herpesvirus 8-Induced Phosphatidylinositol 3-Kinase and RhoA-GTPases Critical for Microtubule Dynamics and Nuclear Delivery of Viral DNA but Dispensable for Binding and Entry. J. Virol. 81: 7941-7959 [Abstract] [Full Text]  
  • Ning, Y., Buranda, T., Hudson, L. G. (2007). Activated Epidermal Growth Factor Receptor Induces Integrin {alpha}2 Internalization via Caveolae/Raft-dependent Endocytic Pathway. J. Biol. Chem. 282: 6380-6387 [Abstract] [Full Text]  
  • Michihara, A., Toda, K., Suenobu, M., Akasaki, K., Tsuji, H. (2007). Decrease of Cholesterol in Mouse Melanoma Causes Secretion of Lysosomal Enzymes. J Biochem 141: 239-250 [Abstract] [Full Text]  
  • Long, G., Pan, X., Kormelink, R., Vlak, J. M. (2006). Functional entry of baculovirus into insect and Mammalian cells is dependent on clathrin-mediated endocytosis.. J. Virol. 80: 8830-8833 [Abstract] [Full Text]  
  • Strunze, S., Trotman, L. C., Boucke, K., Greber, U. F. (2005). Nuclear Targeting of Adenovirus Type 2 Requires CRM1-mediated Nuclear Export. Mol. Biol. Cell 16: 2999-3009 [Abstract] [Full Text]  


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