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Journal of Virology, February 2001, p. 1958-1967, Vol. 75, No. 4
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.4.1958-1967.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Entry of Human Parechovirus 1

Päivi Joki-Korpela,1,2,* Varpu Marjomäki,3 Camilla Krogerus,1 Jyrki Heino,3 and Timo Hyypiä1

Haartman Institute, Department of Virology, University of Helsinki, FIN-00014 Helsinki,1 Department of Virology and MediCity Research Laboratories, University of Turku, FIN-20520 Turku,2 and Department of Biological and Environmental Science, University of Jyväskylä, FIN-40351 Jyväskylä,3 Finland

Received 18 July 2000/Accepted 14 November 2000

Human parechovirus 1 (HPEV-1) is a prototype member of parechoviruses, a recently established picornavirus genus. Although there is preliminary evidence that HPEV-1 recognizes alpha V integrins as cellular receptors, our understanding of early events during HPEV-1 infection is still very limited. The aim of this study was to clarify the entry mechanisms of HPEV-1, including the attachment of the virus onto the host cell surface and subsequent internalization. In blocking experiments with monoclonal antibodies against different receptor candidates, antibodies against alpha V and beta 3 integrin subunits, in particular in combination, appeared to be the most efficient ones in preventing the HPEV-1 infection. To find out whether HPEV-1 uses clathrin-coated vesicles or other routes for the entry into the host cell, we carried out double-labeling experiments of virus-infected cells with anti-HPEV-1 antibodies and antibodies against known markers of the clathrin and the caveolin routes. At the early phase of infection (5 min postinfection [p.i.]) HPEV-1 colocalized with EEA1 (early endosomes), and later, after 30 min p.i., it colocalized with mannose-6-phosphate receptor (late endosomes), whereas no colocalization with caveolin-1 was observed. The data indicate that HPEV-1 utilizes the clathrin-dependent endocytic pathway for entry into the host cells. Interestingly, endocytosed HPEV-1 capsid proteins were observed in the endoplasmic reticulum and cis-Golgi network 30 to 60 min p.i. Depolymerization of microtubules with nocodazole inhibited translocation of the virus to the late endosomes but did not block HPEV-1 replication, suggesting that the RNA genome may be released early during the entry process.

* Corresponding author. Mailing address: Haartman Institute, Department of Virology, P.O. Box 21, FIN-00014 University of Helsinki, Finland. Phone: 358-9-1912 6466. Fax: 358-9-1912 6491. E-mail: paivi.joki-korpela{at}helsinki.fi.

Journal of Virology, February 2001, p. 1958-1967, Vol. 75, No. 4
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.4.1958-1967.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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