title: Identification of the sites of interaction between the scaffold and outer shell in herpes simplex virus-1 capsids by difference electron imaging.
abstract: Formation of herpes simplex virus-1 capsids requires the presence of intact scaffolding proteins. The C terminus of the abundant scaffolding protein associates with the major capsid shell protein VP5 through hydrophobic interactions. After cleavage by the viral encoded protease, which removes their C-terminal 25 aa, the scaffolding proteins are released from the capsid. We have used electron cryomicroscopy and computer image processing to determine, to 13 A, the three-dimensional structures of capsids containing either cleaved or uncleaved scaffolding proteins. Detailed comparisons show that the structures of the outer icosahedral shells are almost identical in the two capsid types. Differences are apparent in the radial distribution of the density inside the capsid shell (within a radius of 460 ) which represents the scaffolding core. However, in both capsid types, the bulk of this internal density exhibits no icosahedral symmetry. Close examination revealed localized regions of icosahedrally arranged extra density at the interface between the outer shell and the scaffold of protease-minus capsids. Rod-like densities extending inwards for approximately 40 from the capsid shell are present under four of the six quasi-equivalent triplex positions. Under triplexes Tb, Tc, and Te, the major additional densities appear as pairs with the rods in each pair situated 37 apart. We propose that these rods are formed by the C-termini of the scaffolding proteins and represent the sites of interaction between the capsid shell and scaffold.
authors.affiliation: Department of Pathology and Laboratory Medicine, University of Texas-Houston Medical School, Houston, TX 77030, USA.
authors.forename: Z H
authors.initials: ZH
authors.lastname: Zhou
authors.forename: S J
authors.initials: SJ
authors.lastname: Macnab
authors.forename: J
authors.initials: J
authors.lastname: Jakana
authors.forename: L R
authors.initials: LR
authors.lastname: Scott
authors.forename: W
authors.initials: W
authors.lastname: Chiu
authors.forename: F J
authors.initials: FJ
authors.lastname: Rixon
mesh_terms: D002213:Capsid / Q000172:deficiency / Q000648:ultrastructure*; D015925:Cryopreservation; D018259:Herpesvirus 1, Human / Q000254:growth & development / Q000648:ultrastructure*; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008854:Microscopy, Electron; D008961:Models, Structural; D012697:Serine Endopeptidases / Q000172:deficiency; D014764:Viral Proteins
chemical_list: D014764:Viral Proteins; C079287:scaffold protein, Herpes simplex virus-1; D012697:Serine Endopeptidases