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Molecular and Cellular Biology, July 2000, p. 4922-4931, Vol. 20, No. 13
Department of Cell Biology, Yale University,
New Haven, Connecticut,1 and
Institute of Biochemistry, ETH-Zürich, Zürich,
Switzerland2
Received 1 November 1999/Returned for modification 8 December
1999/Accepted 4 April 2000
During entry, herpes simplex virus type 1 (HSV-1) releases its
capsid and the tegument proteins into the cytosol of a host cell by
fusing with the plasma membrane. The capsid is then transported to the
nucleus, where it docks at the nuclear pore complexes (NPCs), and the
viral genome is rapidly released into the nucleoplasm. In this study,
capsid association with NPCs and uncoating of the viral DNA were
reconstituted in vitro. Isolated capsids prepared from virus were
incubated with cytosol and purified nuclei. They were found to bind to
the nuclear pores. Binding could be inhibited by pretreating the nuclei
with wheat germ agglutinin, anti-NPC antibodies, or antibodies against
importin
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Herpes Simplex Virus Type 1 Entry into Host Cells:
Reconstitution of Capsid Binding and Uncoating at the Nuclear Pore
Complex In Vitro

. Furthermore, in the absence of cytosol, purified importin
was both sufficient and necessary to support efficient capsid
binding to nuclei. Up to 60 to 70% of capsids interacting with rat
liver nuclei in vitro released their DNA if cytosol and metabolic
energy were supplied. Interaction of the capsid with the nuclear pore
thus seemed to trigger the release of the viral genome, implying that
components of the NPC play an active role in the nuclear events during
HSV-1 entry into host cells.
*
Corresponding author. Mailing address: Haartman
Institute, P. O. Box 21, FIN-00014 University of Helsinki,
Finland. Phone: 358-9-191 26439. Fax: 358-9-191 26700. E-mail:
Paivi.Ojala{at}helsinki.fi.
Present address: Zentrum Biochemie, Medizinische Hochschule,
Hannover, Germany.
Present address: Institute of Biochemistry, ETH-Zürich,
Zürich, Switzerland.
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