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Targeting of Viral Capsids to Nuclear Pores in a Cell‐Free Reconstitution System
Author(s) -
Anderson Fenja,
Savulescu Anca F.,
Rudolph Kathrin,
Schipke Julia,
Cohen Ilana,
Ibiricu Iosune,
Rotem Asaf,
Grünewald Kay,
Sodeik Beate,
Harel Am
Publication year - 2014
Publication title -
traffic
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.677
H-Index - 130
eISSN - 1600-0854
pISSN - 1398-9219
DOI - 10.1111/tra.12209
Subject(s) - viral tegument , capsid , biology , nuclear pore , nucleoplasm , microbiology and biotechnology , nuclear transport , herpes simplex virus , cytoplasm , viral protein , viral replication , virology , nuclear lamina , cell nucleus , virus , nucleolus , nuclear protein , transcription factor , genetics , gene
Many viruses deliver their genomes into the nucleoplasm for viral transcription and replication. Here, we describe a novel cell‐free system to elucidate specific interactions between viruses and nuclear pore complexes ( NPCs ). Nuclei reconstituted in vitro from egg extracts of Xenopus laevis , an established biochemical system to decipher nuclear functions, were incubated with GFP ‐tagged capsids of herpes simplex virus, an alphaherpesvirus replicating in the nucleus. Capsid binding to NPCs was analyzed using fluorescence and field emission scanning electron microscopy. Tegument‐free capsids or viral capsids exposing inner tegument proteins on their surface bound to nuclei, while capsids inactivated by a high‐salt treatment or covered by inner and outer tegument showed less binding. There was little binding of the four different capsid types to nuclei lacking functional NPCs . This novel approach provides a powerful system to elucidate the molecular mechanisms that enable viral structures to engage with NPCs . Furthermore, this assay could be expanded to identify molecular cues triggering viral genome uncoating and nuclear import of viral genomes.