In:
Journal of Virology, American Society for Microbiology, Vol. 80, No. 19 ( 2006-10), p. 9741-9753
Abstract:
Newly assembled herpesvirus capsids travel from the nucleus to the plasma
membrane by a mechanism that is poorly understood. Furthermore, the contribution of cellular proteins to this egress has yet to be
clarified. To address these issues, an in vitro nuclear egress assay that reproduces the exit of herpes simplex virus type 1 (HSV-1) capsids
from nuclei isolated from infected cells was established. As expected, the assay has all the hallmarks of intracellular transport assays,
namely, a dependence on time, energy, and temperature. Surprisingly, it is also dependent on cytosol and was slightly enhanced by infected
cytosol, suggesting an implication of both host and viral proteins in the process. The capsids escaped these nuclei by budding through the
inner nuclear membrane, accumulated as enveloped capsids between the two nuclear membranes, and were released in cytosol exclusively as
naked capsids, exactly as in intact cells. This is most consistent with the view that the virus escapes by crossing the two nuclear membranes
rather than through nuclear pores. Unexpectedly, nuclei isolated at the nonpermissive temperature from cells infected with a U L 26
thermosensitive protease mutant (V701) supported capsid egress. Although electron microscopy, biochemical, and PCR analyses hinted at a
likely reconstitution of capsid maturation, DNA encapsidation could not be confirmed by a traditional SQ test. This assay should prove very
useful for identification of the molecular players involved in HSV-1 nuclear
egress.
Type of Medium:
Online Resource
ISSN:
0022-538X
,
1098-5514
DOI:
10.1128/JVI.00061-06
Language:
English
Publisher:
American Society for Microbiology
Publication Date:
2006
detail.hit.zdb_id:
1495529-5
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