Vasicular stomatitis virus (VSV) is a simple enveloped virus with negative strand RNA genome. It is easily propagated in high yield in tissue culture. Its 5 genes code for 5 proteins whose functions are crudely understood. Temperature sensitive strains with mutations in each gene have been characterized. These properties permit several aspects of VSV infection to be studied at a detailed molecular level. The steps in VSV uncoating that occur after fusion of the viral envelope with the endosomal membrane, and lead to initiation of primary transcription will be defined. Semi-intact cells will be used for this purpose, since the cytoplasmic membrane surfaces are accessible in these cells. M protein- membrane interactions will be studied using hydrophobic photolabeling reagents, in order to identify the specific M protein sequence(s) that interact with viral or cellular membranes during viral budding. The kinetics of VSV assembly from pre-formed elements at the plasma membrane will be determined, as well as any requirements for budding, such as host cell (cytosolic) factors, ATP, transmembrane electrical potentials or specific ion gradients. The transcriptional role of phosphorylation of viral NS protein will be characterized by confirming our preliminary finding that protein kinase activity can be essentially completely separated from the transcriptional complex without loss of transcription activity. The separated kinases will then be run backwards (using ADP as substrate) to effect specific dephosphorylation of NS, which will be correlated with transcriptional activity. The initiation reaction of VSV transcription will be characterized using specific dinucleotide inhibitors and primers, and the initiation site identified by affinity labeling.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI013002-18
Application #
2059943
Study Section
Virology Study Section (VR)
Project Start
1976-09-01
Project End
1995-02-28
Budget Start
1993-08-01
Budget End
1995-02-28
Support Year
18
Fiscal Year
1993
Total Cost
Indirect Cost
Name
University of Medicine & Dentistry of NJ
Department
Physiology
Type
Schools of Medicine
DUNS #
622146454
City
Piscataway
State
NJ
Country
United States
Zip Code
08854
Gao, Y; Lenard, J (1995) Multimerization and transcriptional activation of the phosphoprotein (P) of vesicular stomatitis virus by casein kinase-II. EMBO J 14:1240-7
Lenard, J; Rabson, A; Vanderoef, R (1993) Photodynamic inactivation of infectivity of human immunodeficiency virus and other enveloped viruses using hypericin and rose bengal: inhibition of fusion and syncytia formation. Proc Natl Acad Sci U S A 90:158-62
Stevenson, N R; Lenard, J (1993) Antiretroviral activities of hypericin and rose bengal: photodynamic effects on Friend leukemia virus infection of mice. Antiviral Res 21:119-27
Lenard, J; Vanderoef, R (1993) Photoinactivation of influenza virus fusion and infectivity by rose bengal. Photochem Photobiol 58:527-31
Rigaut, K D; Birk, D E; Lenard, J (1991) Intracellular distribution of input vesicular stomatitis virus proteins after uncoating. J Virol 65:2622-8
Massey, D M; Deans, N; Lenard, J (1990) Phosphorylation of NS protein by vesicular stomatitis virus nucleocapsids: lack of effect during RNA synthesis and separation of kinase from L protein. J Virol 64:3259-64
Subramanian, M; Kovacs, T; Lesiak, K et al. (1990) Inhibition of the RNA polymerase of vesicular stomatitis virus by ppp5'A2'p5'A and related compounds. Antiviral Res 13:81-9
McKenzie, M A; Fawell, S E; Cha, M et al. (1988) Effects of mammalian insulin on metabolism, growth, and morphology of a wall-less strain of Neurospora crassa. Endocrinology 122:511-7
Fawell, S E; McKenzie, M A; Greenfield, N J et al. (1988) Stimulation by mammalian insulin of glycogen metabolism in a wall-less strain of Neurospora crassa. Endocrinology 122:518-23
Bundo-Morita, K; Gibson, S; Lenard, J (1988) Radiation inactivation analysis of fusion and hemolysis by vesicular stomatitis virus. Virology 163:622-4

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