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J Virol, June 1998, p. 4560-4570, Vol. 72, No. 6
Department of Molecular Biology, Princeton
University, Princeton, New Jersey 08544
Received 21 November 1997/Accepted 11 February 1998
The Us9 gene is highly conserved among the alphaherpesviruses
sequenced to date, yet its function remains unknown. In this report, we
demonstrate that the pseudorabies virus (PRV) Us9 protein is present in
infected cell lysates as several phosphorylated polypeptides ranging
from 17 to 20 kDa. Synthesis is first detected at 6 h
postinfection and is sensitive to the DNA synthesis inhibitor phosphonoacetic acid. Unlike the herpes simplex virus type 1 Us9 homolog, which was reported to be associated with nucleocapsids in the
nuclei of infected cells (M. C. Frame, D. J. McGeoch, F. J. Rixon, A. C. Orr, and H. S. Marsden, Virology
150:321-332, 1986), PRV Us9 localizes to the secretory pathway
(predominately to the Golgi apparatus) and not to the nucleus. By
fusing the enhanced green fluorescent protein (EGFP) reporter molecule
to the carboxy terminus of Us9, we demonstrated that Us9 not only is
capable of targeting a Us9-EGFP fusion protein to the Golgi compartment
but also is able to direct efficient incorporation of such chimeric
molecules into infectious viral particles. Moreover, through protease
digestion experiments with Us9-EGFP-containing viral particles, we
demonstrated that the Us9 protein is inserted into the viral envelope
as a type II, tail-anchored membrane protein.
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
The Us9 Gene Product of Pseudorabies Virus, an
Alphaherpesvirus, Is a Phosphorylated, Tail-Anchored Type II
Membrane Protein
*
Corresponding author. Mailing address: Department of
Molecular Biology, Princeton University, Princeton, NJ 08544. Phone: (609) 258-2415. Fax: (609) 258-1035. E-mail:
Lenquist{at}molbiol.princeton.edu.
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