title: Glycoprotein D of herpes simplex virus (HSV) binds directly to HVEM, a member of the tumor necrosis factor receptor superfamily and a mediator of HSV entry.
abstract: Glycoprotein D (gD) is a structural component of the herpes simplex virus (HSV) envelope which is essential for virus entry into host cells. Chinese hamster ovary (CHO-K1) cells are one of the few cell types which are nonpermissive for the entry of many HSV strains. However, when these cells are transformed with the gene for the herpesvirus entry mediator (HVEM), the resulting cells, CHO-HVEM12, are permissive for many HSV strains, such as HSV-1(KOS). By virtue of its four cysteine-rich pseudorepeats, HVEM is a member of the tumor necrosis factor receptor superfamily of proteins. Recombinant forms of gD and HVEM, gD-1(306t) and HVEM(200t), respectively, were used to demonstrate a specific physical interaction between these two proteins. This interaction was dependent on native gD conformation but independent of its N-linked oligosaccharides, as expected from previous structure-function studies. Recombinant forms of gD derived from HSV-1(KOS)rid1 and HSV-1(ANG) did not bind to HVEM(200t), explaining the inability of these viruses to infect CHO-HVEM12 cells. A variant gD protein, gD-1(delta290-299t), showed enhanced binding to HVEM(200t) relative to the binding of gD-1(306t). Competition studies showed that gD-1(delta290-299t) and gD-1(306t) bound to the same region of HVEM(200t), suggesting that the differences in binding to HVEM are due to differences in affinity. These differences were also reflected in the ability of gD-1(delta290-299t) but not gD-1(306t) to block HSV type 1 infection of CHO-HVEM12 cells. By gel filtration chromatography, the complex between gD-1(delta290-299t) and HVEM(200t) had a molecular mass of 113 kDa and a molar ratio of 1:2. We conclude that HVEM interacts directly with gD, suggesting that HVEM is a receptor for virion gD and that the interaction between these proteins is a step in HSV entry into HVEM-expressing cells.
authors.affiliation: School of Dental Medicine, Center for Oral Health Research, University of Pennsylvania, Philadelphia 19104, USA.
authors.forename: J C
authors.initials: JC
authors.lastname: Whitbeck
authors.forename: C
authors.initials: C
authors.lastname: Peng
authors.forename: H
authors.initials: H
authors.lastname: Lou
authors.forename: R
authors.initials: R
authors.lastname: Xu
authors.forename: S H
authors.initials: SH
authors.lastname: Willis
authors.forename: M
authors.initials: M
authors.lastname: Ponce de Leon
authors.forename: T
authors.initials: T
authors.lastname: Peng
authors.forename: A V
authors.initials: AV
authors.lastname: Nicola
authors.forename: R I
authors.initials: RI
authors.lastname: Montgomery
authors.forename: M S
authors.initials: MS
authors.lastname: Warner
authors.forename: A M
authors.initials: AM
authors.lastname: Soulika
authors.forename: L A
authors.initials: LA
authors.lastname: Spruce
authors.forename: W T
authors.initials: WT
authors.lastname: Moore
authors.forename: J D
authors.initials: JD
authors.lastname: Lambris
authors.forename: P G
authors.initials: PG
authors.lastname: Spear
authors.forename: G H
authors.initials: GH
authors.lastname: Cohen
authors.forename: R J
authors.initials: RJ
authors.lastname: Eisenberg
mesh_terms: D000818:Animals; D016466:CHO Cells; D002522:Chlorocebus aethiops; D002850:Chromatography, Gel; D006224:Cricetinae; D011487:Protein Conformation; D011817:Rabbits; D018124:Receptors, Tumor Necrosis Factor / Q000378:metabolism*; D053240:Receptors, Tumor Necrosis Factor, Member 14; D011991:Receptors, Virus / Q000378:metabolism*; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D018124:Receptors, Tumor Necrosis Factor; D053240:Receptors, Tumor Necrosis Factor, Member 14; D011991:Receptors, Virus; C506028:TNFRSF14 protein, human; D014759:Viral Envelope Proteins; C039447:glycoprotein D, Human herpesvirus 1