Roles of triplex and scaffolding proteins in herpes simplex virus type 1 capsid formation suggested by structures of recombinant particles.
Typical herpes simplex virus (HSV) capsids contain seven proteins that form a T=16 icosahedron of 1,250-A diameter. Infection of cells with recombinant baculoviruses expressing two of these proteins, VP5 (which forms the pentons and hexons in typical HSV capsids) and VP19C (a component of the triplexes that connect adjacent capsomeres), results in the formation of spherical particles of 880-A diameter. Electron cryomicroscopy and computer reconstruction revealed that these particles possess a T=7 icosahedral symmetry, having 12 pentons and 60 hexons. Among the characteristic structural features of the particle are the skewed appearance of the hexons and the presence of intercapsomeric mass densities connecting the middle domain of one hexon subunit to the lower domain of a subunit in the adjacent hexon. We interpret these connecting masses as being formed by VP19C. Comparison of the connecting masses with the triplexes, which occupy equivalent positions in the T=16 capsid, reveals the probable locations of the single VP19C and two VP23 molecules that make up the triplex. Their arrangement suggests that the two triplex proteins have different roles in controlling intercapsomeric interactions and capsid stability. The nature of these particles and of other aberrant forms made in the absence of scaffold demonstrates the conformational adaptability of the capsid proteins and illustrates how VP23 and the scaffolding protein modulate the nature of the VP5-VP19C network to ensure assembly of the functional T=16 capsid.
Saad
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A
Verna and Marrs McLean Department of Biochemistry, Baylor College of Medicine, Houston, Texas 77030, USA.
Zhou
Z H
ZH
Jakana
J
J
Chiu
W
W
Rixon
F J
FJ
D002213:Capsid / Q000096:biosynthesis / Q000235:genetics / Q000648:ultrastructure*; D036022:Capsid Proteins; D020285:Cryoelectron Microscopy; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000648:ultrastructure*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000648:ultrastructure; D011995:Recombination, Genetic; D014771:Virion / Q000648:ultrastructure
D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; C082563:VP19 protein, Human herpesvirus 1; C079750:VP23 protein, Human herpesvirus 1; C442523:VP5 protein, Herpes simplex virus type 1