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Journal of Virology, July 2009, p. 6335-6346, Vol. 83, No. 13
0022-538X/09/$08.00+0 doi:10.1128/JVI.00375-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Ling Li,
,
and
Rozanne M. Sandri-Goldin*
Department of Microbiology and Molecular Genetics, School of Medicine, University of California, Irvine, California 92697-4025
Received 19 February 2009/ Accepted 7 April 2009
Herpes simplex virus 1 (HSV-1) protein ICP27 has been shown to shuttle between the nucleus and cytoplasm and to bind viral RNA during infection. ICP27 was found to interact with the cellular RNA export adaptor protein Aly/REF, which is part of the TREX complex, and to relocalize Aly/REF to viral replication sites. ICP27 is exported to the cytoplasm through the export receptor TAP/NXF1, and ICP27 must be able to interact with TAP/NXF1 for efficient export of HSV-1 early and late transcripts. We examined the dynamics of ICP27 movement and its localization with respect to Aly/REF and TAP/NXF1 in living cells during viral infection. Recombinant viruses with a yellow fluorescent protein (YFP) tag on the N or C terminus of ICP27 were constructed. While the N-terminally tagged ICP27 virus behaved like wild-type HSV-1, the C-terminally tagged virus was defective in viral replication and gene expression, and ICP27 was confined to the nucleus, suggesting that the C-terminal YFP tag interfered with ICP27's C-terminal interactions, including the interaction with TAP/NXF1. To assess the role of Aly/REF and TAP/NXF1 in viral RNA export, these factors were knocked down using small interfering RNA. Knockdown of Aly/REF had little effect on the export of ICP27 or poly(A)+ RNA during infection. In contrast, a decrease in TAP/NXF1 levels severely impaired export of ICP27 and poly(A)+ RNA. We conclude that TAP/NXF1 is essential for ICP27-mediated export of RNA during HSV-1 infection, whereas Aly/REF may be dispensable.
Published ahead of print on 15 April 2009.
L.A.J. and L.L. contributed equally to this work.
Present address: Verdezyne, Inc., Carlsbad, CA.
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