ATP-Dependent localization of the herpes simplex virus capsid protein VP26 to sites of procapsid maturation.
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.
Chi
J H
JH
Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, Bronx, New York, New York 10461, USA.
Wilson
D W
DW
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D019556:COS Cells; D002213:Capsid / Q000235:genetics* / Q000378:metabolism*; D036022:Capsid Proteins; D049452:Green Fluorescent Proteins; D018259:Herpesvirus 1, Human / Q000378:metabolism*; D007150:Immunohistochemistry; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D013696:Temperature; D019065:Virus Assembly
D036022:Capsid Proteins; D008164:Luminescent Proteins; C077409:capsid protein VP26, herpes simplex virus type 1; C018315:hexon capsid protein, Adenovirus; D049452:Green Fluorescent Proteins; D000255:Adenosine Triphosphate