The long term goal of the studies described in this proposal is to develop methods by which retroviruses and retroviral vectors may be targeted to specific cell types. These investigations are of medical significance because of the potential therapeutic value of employing targeted retroviral vectors for the treatment of genetically determined birth defects. Moreover, these studies should yield detailed information as to the mechanisms by which the Moloney murine leukemia virus (Mo-MuLV) envelope (env) proteins become incorporated into viral particles, the structure/function relationships of the Mo-MuLV env proteins, and the putative association of the env proteins with the Mo-MuLV core (gag) proteins. The model system described is ideal for the elucidation of general mechanisms of viral assembly and of cell surface and cytoplasmic protein interaction.
The specific aims are as follows: 1. We will undertake studies of mutant Mo-MuLV env proteins (the gp70/p15E complex) with respect to their subcellular localization, ability to be incorporated into viral particles, and facilitation of viral entry into cells. 2. Non-retroviral """"""""targeting"""""""" proteins, or fusions of such proteins with the Mo-MuLV envelope proteins will be examined to determine the factors which permit incorporation of heterologous proteins into retroviral particles. 3. Mutations in Mo-MuLV gag genes which may affect the incorporation of env proteins into virions will be generated and analyzed. Gag gene sequences responsible for directing the Pr65gag polyprotein to cell membranes and viral particles will be identified by analysis of gag gene fusions to indicator genes.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA047088-03
Application #
3190565
Study Section
Experimental Virology Study Section (EVR)
Project Start
1988-05-01
Project End
1991-04-30
Budget Start
1990-05-01
Budget End
1991-04-30
Support Year
3
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Oregon Health and Science University
Department
Type
Schools of Medicine
DUNS #
009584210
City
Portland
State
OR
Country
United States
Zip Code
97239
Clish, C B; Peyton, D H; Barklis, E (1998) Solution structures of human immunodeficiency virus type 1 (HIV-1) and moloney murine leukemia virus (MoMLV) capsid protein major-homology-region peptide analogs by NMR spectroscopy. Eur J Biochem 257:69-77
Zhang, Y; Qian, H; Love, Z et al. (1998) Analysis of the assembly function of the human immunodeficiency virus type 1 gag protein nucleocapsid domain. J Virol 72:1782-9
Barklis, E; McDermott, J; Wilkens, S et al. (1998) Organization of HIV-1 capsid proteins on a lipid monolayer. J Biol Chem 273:7177-80
Barklis, E; McDermott, J; Wilkens, S et al. (1997) Structural analysis of membrane-bound retrovirus capsid proteins. EMBO J 16:1199-213
Zhang, Y; Barklis, E (1997) Effects of nucleocapsid mutations on human immunodeficiency virus assembly and RNA encapsidation. J Virol 71:6765-76
Birrell, G B; Hedberg, K K; Barklis, E et al. (1997) Partial isolation from intact cells of a cell surface-exposed lysophosphatidylinositol-phospholipase C. J Cell Biochem 65:550-64
McDermott, J; Farrell, L; Ross, R et al. (1996) Structural analysis of human immunodeficiency virus type 1 Gag protein interactions, using cysteine-specific reagents. J Virol 70:5106-14
Hansen, M S; Barklis, E (1995) Structural interactions between retroviral Gag proteins examined by cysteine cross-linking. J Virol 69:1150-9
Wang, C T; Barklis, E (1993) Assembly, processing, and infectivity of human immunodeficiency virus type 1 gag mutants. J Virol 67:4264-73
Wang, C T; Zhang, Y; McDermott, J et al. (1993) Conditional infectivity of a human immunodeficiency virus matrix domain deletion mutant. J Virol 67:7067-76

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