title: ICAM-1 Binding Rhinoviruses Enter HeLa Cells via Multiple Pathways and Travel to Distinct Intracellular Compartments for Uncoating.
abstract: Of the more than 150 human rhinovirus (RV) serotypes, 89 utilize intercellular adhesion molecule-1 (ICAM-1) for cell entry. These belong either to species A or B. We recently demonstrated that RV-B14 and RV-A89, despite binding this same receptor, are routed into distinct endosomal compartments for release of their RNA into the cytosol. To gain insight into the underlying mechanism we now comparatively investigate the port of entry, temperature-dependence of uncoating, and intracellular routing of RV-B3, RV-B14, RV-A16, and RV-A89 in HeLa cells. The effect of various drugs blocking distinct stages on the individual pathways was determined via comparing the number of infected cells in a TissueFaxs instrument. We found that RV-B14 and RV-A89 enter via clathrin-, dynamin-, and cholesterol-dependent pathways, as well as by macropinocytosis. Drugs interfering with actin function similarly blocked entry of all four viruses, indicating their dependence on a dynamic actin network. However, uniquely, RV-A89 was able to produce progeny when internalized at 20 °C followed by neutralizing the endosomal pH and further incubation at 37 °C. Blocking dynein-dependent endosomal transport prevented uncoating of RV-A16 and RV-A89, but not of RV-B3 and RV-B14, indicative for routing of RV-A16 and RV-A89 into the endocytic recycling compartment for uncoating. Our results call for caution when developing drugs aimed at targeting entry or intracellular trafficking of all rhinovirus serotypes.
authors.affiliation: Department of Pathophysiology and Allergy Research, Medical University of Vienna, Währinger Gürtel 18-20, A-1090 Wien, Austria. halehganjian85@gmail.com.
authors.forename: Haleh
authors.initials: H
authors.lastname: Ganjian
authors.affiliation: Department of Pathophysiology and Allergy Research, Medical University of Vienna, Währinger Gürtel 18-20, A-1090 Wien, Austria. a1368365@unet.univie.ac.at.
authors.forename: Christin
authors.initials: C
authors.lastname: Zietz
authors.affiliation: Department of Pathophysiology and Allergy Research, Medical University of Vienna, Währinger Gürtel 18-20, A-1090 Wien, Austria. diana.mechtcheriakova@meduniwien.ac.at.
authors.forename: Diana
authors.initials: D
authors.lastname: Mechtcheriakova
authors.affiliation: Max F. Perutz Laboratories, Department of Medical Biochemistry, Medical University of Vienna, Dr. Bohr Gasse 9/3, A-1030 Vienna, Austria. dieter.blaas@meduniwien.ac.at.
authors.forename: Dieter
authors.initials: D
authors.lastname: Blaas
authors.affiliation: Department of Pathophysiology and Allergy Research, Medical University of Vienna, Währinger Gürtel 18-20, A-1090 Wien, Austria. renate.fuchs@meduniwien.ac.at.
authors.forename: Renate
authors.initials: R
authors.lastname: Fuchs
mesh_terms: D001692:Biological Transport; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism*; D011991:Receptors, Virus / Q000378:metabolism*; D012229:Rhinovirus / Q000502:physiology*; D013696:Temperature; D053585:Virus Attachment; D053586:Virus Internalization; D057646:Virus Uncoating
chemical_list: D011991:Receptors, Virus; D018799:Intercellular Adhesion Molecule-1
keywords: ICAM-1; actin; clathrin; dynamin; dynein; endocytosis; human rhinoviruses; macropinocytosis; productive uncoating