The overall objective of the proposed research program is to define the synthesis, processing and localization of the herpes simplex virus (HSV)-specific glycoproteins associated with the virus-infected cell. The proposed studies may be separated into three major areas of focus. The first aspect concerns defining the mechanisms responsible for the targeting and retention of HSV glycoproteins within the nucleus, the site of nucleocapsid envelopment. In an effort to identify regions of gB and gC required for nuclear association and retention, chimeric genes between VSV-G and HSV glycoprotein genes will be constructed. In addition, deletions in glycoprotein genes will be generated to analyze their effect on the subcellular trafficking of these proteins. Experiments will also be conducted to identify and isolate proteins which interact with and possibly retain the HSV glycoproteins in the nucleus. Approaches will include, immunoprecipitation of nuclear associated glycoproteins, cross- linking analysis and affinity chromatography. The second aspect of the study will focus on the use of bifunctional chemical cross- linking reagents to define the organization and intermolecular interactions of the HSV glycoproteins present on the viral envelope and the plasma membrane of virus-infected cells. Productively-infected cells as well as cells transfected with cloned glycoprotein genes will be used in this aspect of the study. The third aspect of the study will focus on the synthesis, processing and functional role of the HSV-2 glycoprotein gG-2. Experiments will be conducted to define the cleavage event associated with the precursor form of the glycoprotein. Studies will include defining the site of the cleavage on the precursor and to determine where in the infected cell the cleavage event occurs. Finally, experiments are proposed to determine the role the secreted cleavage product (34K) may play in the induction of the humoral immune response following an HSV-2 infection.
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