The overall goal of the proposed research program is to define the mechanism by which virion glycoproteins of HSV mediate virus entry into neuronal and non-neuronal cells. After the initial interaction virion gC and host cell heparin sulfate, four glycoproteins, gB, gD and gH/gL complex mediate virus-cell fusion at the plasma membrane. A major function gD is to interact with a specific cellular receptor which apparently triggers later events involving gB and gH/gL. A recent finding from Dr. Patricia Spear's laboratory (Northwestern University has shown that a protein designated HVEM (herpes virus entry mediator), when expressed by normally non-permissive cells, makes those cells permissive for HSV-1 entry. HVEM is a member of the tumor necrosis factor receptor protein family. The co-principal investigators, Drs. Eisenberg and Cohen, have initiated a collaboration with Dr. Spear to study the interaction of gD and HVEM. Drs. Eisenberg and Cohen have subcloned a truncated form of the HVEM gene (HVEMt) into the baculovirus expression system and found that purified truncated gD (gDt) binds directly to purified HVEMt in vitro. Thus, the two proteins can interact directly, without any requirement for another viral or cellular protein. They propose to extend these initial observations and to further detail the stoichiometry and affinity of this interaction, as well as to attempt co-crystallization. gD and HVEM mutants will be used to localize the binding sites and to measure the effect of mutation on the affinity of the interaction. They will explore how the interaction between gD and HVEM triggers the next steps of virus entry involving other viral and/or possibly other cellular molecules. This will be done both in vitro with purified glycoproteins, and in vivo, with virions and cells. Preliminary data suggest that mutations in gD can either enhance or ablate binding to HVEM. It is possible that gD interacts with more than one receptor, depending on the cell type and differences in gD structure. They will explore whether strain differences in gD affect viral tropism, especially in viral isolates from encephalitis patients versus isolates from other sites in the human host.
Three specific aims are proposed: (1) to characterize the interaction between gD and HVEM in vitro; (2) to localize domains involved in the GD-HVEM interaction; and (3) to examine the consequences of gD-receptor interactions for virus entry into neuronal and non-neuronal cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
3R01NS036731-03S1
Application #
6320304
Study Section
Virology Study Section (VR)
Program Officer
Kerza-Kwiatecki, a P
Project Start
1997-07-15
Project End
2002-06-30
Budget Start
1999-07-01
Budget End
2000-06-30
Support Year
3
Fiscal Year
2000
Total Cost
$50,000
Indirect Cost
Name
University of Pennsylvania
Department
Pathology
Type
Schools of Veterinary Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Atanasiu, Doina; Cairns, Tina M; Whitbeck, J Charles et al. (2013) Regulation of herpes simplex virus gB-induced cell-cell fusion by mutant forms of gH/gL in the absence of gD and cellular receptors. MBio 4:
Maurer, Ulrike E; Zeev-Ben-Mordehai, Tzviya; Pandurangan, Arun Prasad et al. (2013) The structure of herpesvirus fusion glycoprotein B-bilayer complex reveals the protein-membrane and lateral protein-protein interaction. Structure 21:1396-405
Krummenacher, Claude; CarfĂ­, Andrea; Eisenberg, Roselyn J et al. (2013) Entry of herpesviruses into cells: the enigma variations. Adv Exp Med Biol 790:178-95
Atanasiu, Doina; Saw, Wan Ting; Gallagher, John R et al. (2013) Dual split protein-based fusion assay reveals that mutations to herpes simplex virus (HSV) glycoprotein gB alter the kinetics of cell-cell fusion induced by HSV entry glycoproteins. J Virol 87:11332-45
Eisenberg, Roselyn J; Atanasiu, Doina; Cairns, Tina M et al. (2012) Herpes virus fusion and entry: a story with many characters. Viruses 4:800-32
Shelly, Spencer S; Cairns, Tina M; Whitbeck, J Charles et al. (2012) The membrane-proximal region (MPR) of herpes simplex virus gB regulates association of the fusion loops with lipid membranes. MBio 3:
Lazear, Eric; Whitbeck, J Charles; Ponce-de-Leon, Manuel et al. (2012) Antibody-induced conformational changes in herpes simplex virus glycoprotein gD reveal new targets for virus neutralization. J Virol 86:1563-76
Bernstein, David I; Earwood, Julie D; Bravo, Fernando J et al. (2011) Effects of herpes simplex virus type 2 glycoprotein vaccines and CLDC adjuvant on genital herpes infection in the guinea pig. Vaccine 29:2071-8
Di Giovine, Paolo; Settembre, Ethan C; Bhargava, Arjun K et al. (2011) Structure of herpes simplex virus glycoprotein D bound to the human receptor nectin-1. PLoS Pathog 7:e1002277
Cairns, Tina M; Whitbeck, J Charles; Lou, Huan et al. (2011) Capturing the herpes simplex virus core fusion complex (gB-gH/gL) in an acidic environment. J Virol 85:6175-84

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