Structure of the herpes simplex virus capsid: effects of extraction with guanidine hydrochloride and partial reconstitution of extracted capsids.
Viral B capsids were purified from cells infected with herpes simplex virus type 1 and extracted in vitro with 2.0 M guanidine hydrochloride (GuHCl). Sodium dodecyl sulfate-polyacrylamide gel analyses demonstrated that extraction resulted in the removal of greater than 95% of capsid proteins VP22a and VP26 while there was only minimal (less than 10%) loss of VP5 (the major capsid protein), VP19, and VP23. Electron microscopic analysis of extracted capsids revealed that the pentons and the material found inside the cavity of B capsids (primarily VP22a) were removed nearly quantitatively, but extracted capsids remained otherwise structurally intact. Few, if any, hexons were lost; the capsid diameter was not greatly affected; and its icosahedral symmetry was still clearly evident. The results demonstrate that neither VP19 nor VP23 could constitute the capsid pentons. Like the hexons, the pentons are most likely composed of VP5. When B capsids were treated with 2.0 M GuHCl and then dialyzed to remove GuHCl, two bands of viral material were separated by sucrose density gradient ultracentrifugation. The more rapidly migrating of the two consisted of capsids which lacked pentons and VP22a but had a full complement of VP26. Thus, VP26 must have reassociated with extracted capsids during dialysis. The more slowly migrating band consisted of torus-shaped structures approximately 60 nm in diameter which were composed entirely of VP22a. These latter structures closely resembled torus-shaped condensates often seen in the cavity of native B capsids. The results suggest a similarity between herpes simplex virus type 1 B capsids and procapsids of Salmonella bacteriophage P22. Both contain an internal protein (VP22a in the case of HSV-1 B capsids and gp8 or "scaffolding" protein in phage P22) that can be extracted in vitro with GuHCl and that is absent from mature virions.
Newcomb
W W
WW
Department of Microbiology, University of Virginia Health Sciences Center, Charlottesville 22908.
Brown
J C
JC
D000818:Animals; D002213:Capsid / Q000302:isolation & purification* / Q000378:metabolism / Q000648:ultrastructure; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D019791:Guanidine; D006146:Guanidines; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008970:Molecular Weight; D018139:Simplexvirus / Q000254:growth & development / Q000378:metabolism / Q000648:ultrastructure*
D006146:Guanidines; D046911:Macromolecular Substances; D019791:Guanidine