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Journal of Virology, February 2000, p. 1468-1476, Vol. 74, No. 3
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
ATP-Dependent Localization of the Herpes Simplex
Virus Capsid Protein VP26 to Sites of Procapsid Maturation
Jung Hee I.
Chi and
Duncan W.
Wilson*
Department of Developmental and Molecular
Biology, Albert Einstein College of Medicine, Bronx, New York, New
York 10461
Received 24 August 1999/Accepted 5 November 1999
The herpes simplex virus type 1 (HSV-1) capsid shell is composed of
four major polypeptides, VP5, VP19c, VP23, and VP26. VP26, a 12-kDa
polypeptide, is associated with the tips of the capsid hexons formed by
VP5. Mature capsids form upon angularization of the shell of
short-lived, fragile spherical precursors termed procapsids. The cold
sensitivity and short-lived nature of the procapsid have made its
isolation and biochemical analysis difficult, and it remains unclear
whether procapsids contain bound VP26 or whether VP26 is recruited
following shell angularization. By indirect immunocytochemical analysis
of virally expressed VP26 and by direct visualization of a transiently
expressed VP26-green fluorescent protein fusion, we show that VP26
fails to specifically localize to intranuclear procapsids accumulated
following incubation of the temperature-sensitive HSV mutant
tsProt.A under nonpermissive conditions. However, following
a downshift to the permissive temperature, which allows procapsid
maturation to proceed, VP26 was seen to concentrate at intranuclear
sites which also contained epitopes specific to mature, angularized
capsids. Like the formation of these epitopes, the association of VP26
with maturing capsids was blocked in a reversible fashion by the
depletion of intracellular ATP. We conclude that unlike the other major
capsid shell proteins, VP26 is recruited in an ATP-dependent fashion
after procapsid maturation begins.
*
Corresponding author. Mailing address: Department of
Developmental and Molecular Biology, Albert Einstein College of
Medicine, 1300 Morris Park Ave., Bronx, New York, NY 10461. Phone:
(718) 430-2305. Fax: (718) 430-8567. E-mail:
wilson{at}aecom.yu.edu.
Journal of Virology, February 2000, p. 1468-1476, Vol. 74, No. 3
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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