Herpes simplex virus (HSV) and other alpha-herpesviruses possess the amazing innate ability to spread very rapidly from infected cells to neighboring cells, especially in biologically important epithelial and neuronal tissues. This process of cell-to-cell spread involves specialized virus machinery that: i) directs virus particles to junctions formed between cells and ii) allows the virus to cross cell junctions and enter neighboring cells. HSV glycoprotein gE/gl plays an important role in both these facets of virus cell-to-cell spread but, significantly, does not play a role in entry of extracellular virus particles at the apical surfaces of epithelial cells. As such gE/gl is an important molecular handle to study herpesvirus cell-to-cell spread. The cytoplasmic domain of gE/gl affects sorting of the glycoprotein to trans-Golgi network (TGN) of cells and plays an important role in virus envelopment into cytosolic membranes. The TGN is the major site of basolateral/apical sorting in polarized cells and gE/gl promotes sorting of nascent virions specifically to basolateral surfaces of cells and cell junctions. Moreover, we have two new pieces of evidence that the extracellular domain of gE/gl binds to components of cell junctions, cellular receptors, and that this promotes infection of adjacent cells. First, mutations in the extracellular domain of gE significantly diminish cell-to-cell spread. Second, expression of gE/gl in cells, in trans, interferes with cell-to cell spread. The ? aims of the research proposed here are to: i) better understand the intracellular traffic of gE/gl and how this determines sorting of nascent virions to cell junctions, ii) to characterize the contribution of gE/gl and other HSV glycoproteins in cytoplasmic envelopment of nucleocapsids, iii) to characterize the extracellular domains of gE/gl in mediating movement of viruses across cell junctions and into adjacent cells, and iv) to examine the effects of gE/gl mutations on HSV spread in the cornea. These studies will help elucidate this poorly understood process of herpesvirus cell-to-cell spread, and will also expand our knowledge of how a herpesvirus moves through mucosal epithelium and within the nervous system. ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
9R01AI055051-06
Application #
6612471
Study Section
Virology Study Section (VR)
Program Officer
Beisel, Christopher E
Project Start
1998-07-01
Project End
2008-04-30
Budget Start
2003-05-15
Budget End
2004-04-30
Support Year
6
Fiscal Year
2003
Total Cost
$302,000
Indirect Cost
Name
Oregon Health and Science University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
096997515
City
Portland
State
OR
Country
United States
Zip Code
97239
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Ryckman, Brent J; Chase, Marie C; Johnson, David C (2008) HCMV gH/gL/UL128-131 interferes with virus entry into epithelial cells: evidence for cell type-specific receptors. Proc Natl Acad Sci U S A 105:14118-23
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