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Porcine circovirus 2 infection of epithelial cells is clathrin-, caveolae- and dynamin-independent, actin and Rho-GTPase-mediated, and enhanced by cholesterol depletion.
Virus Res. 2009 Jan; 139(1):1-9.VR

Abstract

Epithelial cells are the major in vivo target cells for porcine circovirus type 2 (PCV2). Although these cells are used for most studies of PCV2 gene expression and, little is known on PCV2 entry, attachment and internalization, in epithelial cells. PCV2 attachment to epithelial cells occurred rapidly and in a time-dependent manner. In contrast to attachment, internalization was slow. Immunofluorescent stainings revealed that during internalization, PCV2 co-localized with clathrin, but not caveolin. Blocking clathrin-mediated endocytosis increased instead of decreased the number of PCV2-infected cells by threefold, suggesting that it does not represent the main internalization pathway leading to a full replication. Further analysis with different inhibitors revealed that also macropinocytosis, dynamin-dependent internalization and membrane cholesterol play no role in PCV2 entry that leads to infection. Inhibition of small GTPases with Clostridium difficile toxin B reduced the number of PCV2-infected PK-15, SK and STs to 63+/-25%, 47+/-21% and 14+/-6%, respectively. Finally, inhibiting actin polymerization also blocked PCV2 infection, showing the need for actin during PCV2 infection. Together, these data indicate that a dynamin- and cholesterol-independent, but actin- and small GTPase-dependent pathway, allows PCV2 internalization in epithelial cells that leads to infection and that clathrin-mediated PCV2 internalization in epithelial cells is not followed by a full replication.

Authors+Show Affiliations

Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18952130

Citation

Misinzo, G, et al. "Porcine Circovirus 2 Infection of Epithelial Cells Is Clathrin-, Caveolae- and Dynamin-independent, Actin and Rho-GTPase-mediated, and Enhanced By Cholesterol Depletion." Virus Research, vol. 139, no. 1, 2009, pp. 1-9.
Misinzo G, Delputte PL, Lefebvre DJ, et al. Porcine circovirus 2 infection of epithelial cells is clathrin-, caveolae- and dynamin-independent, actin and Rho-GTPase-mediated, and enhanced by cholesterol depletion. Virus Res. 2009;139(1):1-9.
Misinzo, G., Delputte, P. L., Lefebvre, D. J., & Nauwynck, H. J. (2009). Porcine circovirus 2 infection of epithelial cells is clathrin-, caveolae- and dynamin-independent, actin and Rho-GTPase-mediated, and enhanced by cholesterol depletion. Virus Research, 139(1), 1-9. https://doi.org/10.1016/j.virusres.2008.09.005
Misinzo G, et al. Porcine Circovirus 2 Infection of Epithelial Cells Is Clathrin-, Caveolae- and Dynamin-independent, Actin and Rho-GTPase-mediated, and Enhanced By Cholesterol Depletion. Virus Res. 2009;139(1):1-9. PubMed PMID: 18952130.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Porcine circovirus 2 infection of epithelial cells is clathrin-, caveolae- and dynamin-independent, actin and Rho-GTPase-mediated, and enhanced by cholesterol depletion. AU - Misinzo,G, AU - Delputte,P L, AU - Lefebvre,D J, AU - Nauwynck,H J, Y1 - 2008/11/12/ PY - 2007/12/13/received PY - 2008/09/10/revised PY - 2008/09/11/accepted PY - 2008/10/28/pubmed PY - 2009/3/18/medline PY - 2008/10/28/entrez SP - 1 EP - 9 JF - Virus research JO - Virus Res. VL - 139 IS - 1 N2 - Epithelial cells are the major in vivo target cells for porcine circovirus type 2 (PCV2). Although these cells are used for most studies of PCV2 gene expression and, little is known on PCV2 entry, attachment and internalization, in epithelial cells. PCV2 attachment to epithelial cells occurred rapidly and in a time-dependent manner. In contrast to attachment, internalization was slow. Immunofluorescent stainings revealed that during internalization, PCV2 co-localized with clathrin, but not caveolin. Blocking clathrin-mediated endocytosis increased instead of decreased the number of PCV2-infected cells by threefold, suggesting that it does not represent the main internalization pathway leading to a full replication. Further analysis with different inhibitors revealed that also macropinocytosis, dynamin-dependent internalization and membrane cholesterol play no role in PCV2 entry that leads to infection. Inhibition of small GTPases with Clostridium difficile toxin B reduced the number of PCV2-infected PK-15, SK and STs to 63+/-25%, 47+/-21% and 14+/-6%, respectively. Finally, inhibiting actin polymerization also blocked PCV2 infection, showing the need for actin during PCV2 infection. Together, these data indicate that a dynamin- and cholesterol-independent, but actin- and small GTPase-dependent pathway, allows PCV2 internalization in epithelial cells that leads to infection and that clathrin-mediated PCV2 internalization in epithelial cells is not followed by a full replication. SN - 0168-1702 UR - https://www.unboundmedicine.com/medline/citation/18952130/Porcine_circovirus_2_infection_of_epithelial_cells_is_clathrin__caveolae__and_dynamin_independent_actin_and_Rho_GTPase_mediated_and_enhanced_by_cholesterol_depletion_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0168-1702(08)00325-0 DB - PRIME DP - Unbound Medicine ER -