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J. Biol. Chem., Vol. 276, Issue 6, 3952-3962, February 9, 2001
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From the Departments of Minor group human rhinoviruses (exemplified by
human rhinovirus serotype 2 (HRV2)) use members of the low density
lipoprotein receptor family for cell entry; all these receptors possess
clathrin-coated pit localization signals. Viral infection should thus
be inhibited under conditions of impaired clathrin-mediated
endocytosis. However, Madshus et al. reported an increase
in the cytopathic effect of HRV2 infection in HEp-2 cells upon
suppression of clathrin-dependent endocytosis by hypotonic
shock and potassium depletion (Madshus, I. H., Sandvig, K.,
Olsnes, S., and van Deurs, B. (1987) J. Cell. Physiol.
131, 14-22.) To resolve this apparent contradiction, we investigated
the binding, internalization, conformational changes, and productive
uncoating of HRV2 in HeLa cells subjected to hypotonic shock and
potassium depletion. This treatment led to an increase in HRV2 binding,
with internalization being barely affected. The generation of
C-antigenic particles requiring pH
Inhibition of Clathrin-dependent Endocytosis Has
Multiple Effects on Human Rhinovirus Serotype 2 Cell Entry*
,,
Pathophysiology and
§ Medical Chemistry, University of Vienna, A-1090 Vienna,
Austria and the Division of Biochemistry, Institute of Medical
Biochemistry, Vienna Biocenter, University of Vienna,
A-1030 Vienna, Austria
5.6 was strongly reduced due to an
elevation of the pH in endosomal compartments. However, K+
depletion only slightly affected de novo viral protein
synthesis, suggesting that productivity of viral RNA in the cytoplasm
is enhanced and thus compensates for the reduction in C-antigenic particles. The distinct steps in the entry pathway of HRV2 are thus
differently influenced by potassium depletion. Viral internalization under conditions of inhibited clathrin-dependent
endocytosis without the need to disturb the ionic milieu was confirmed
in HeLa cells overexpressing the nonfunctional dynamin-1 mutant K44A.
Unexpectedly, overexpression of dynamin-1 K44A resulted in elevated
endosomal pH compared with overexpression of wild-type dynamin.
*
This work was supported by Austrian Science Foundation
Grants P-12967-GEN and P-10618-MED (to R. F.) and P-12269-MOB (to
D. B.) and Jubiläumsfonds der Österreichischen
Nationalbank Grant 7511 (to R. F.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of
Pathophysiology, University of Vienna, Währinger Gürtel
18-20, A-1090 Vienna, Austria. Tel.: 43-1-40-400-5127; Fax:
43-1-40-400-5130; E-mail: renate.fuchs@akh-wien.ac.at.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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