We are interested in gene expression and molecular pathogenesis of hepatitis B virus (HBV). In this proposal we will study: 1) the expression of the surface antigen (HBsAg) gene. The expression of the HBsAg gene is presumably regulated by two different promoters. Deletion mapping experiments will be performed to map these two promoters. Subsequently, the mapped -promoter sequences will be used in gel-retardation assays to characterize the protein factors that regulate these two promoters. 2) the transportation of HBsAg in cells and the assembly of 22nm HBsAg particles. We will mutate and express HBsAg proteins in order to study the functions of various domains of the HBsAg molecule on the transportation of HBsAg proteins in cells and on the assembly of 22nm particles. 3) the translocation of the core antigen (HBcAg) into membranes. The precore region of the HBcAg gene contains a signal peptide which can lead to the secretion of another HBV-related antigen, the e antigen, from cells. HBcAg with the precore region translated in vitro, however, cannot be translocated into microsomal membranes. We will fuse the HBcAg coding sequence to the preprolactin coding sequence and perform in vitro translation experiments to examine whether the precore region contains a defective signal peptide which requires helper factors to translocate HBcAg into membranes or the HBcAg sequence contains signals that inhibit its own translocation into membranes. 4) the nature of the polymerase gene product. The polymerase gene with or without the preceding HBcAg coding sequence will be expressed in mammalian cells. We will examine whether the polymerase is expressed as a HBcAg-pol fusion protein and whether post-translational processing is required for activating this enzyme. 5) the role of cytotoxic T-lymphocytes (CTL) in HBV-induced hepatitis. We will test the cytotoxicity of CTL isolated from chronic HBV carriers on HBV gene-transfected autologous fibroblasts. The goal of this project is to understand the mechanism of HBV-induced hepatitis. 6) the autocrine growth of hepG2 cells, a human hepatoma cell line. We will use biochemical methods to purify and characterize hepG2-derived autocrine factors.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI026244-03
Application #
3139962
Study Section
Experimental Virology Study Section (EVR)
Project Start
1988-05-01
Project End
1992-04-30
Budget Start
1990-05-01
Budget End
1991-04-30
Support Year
3
Fiscal Year
1990
Total Cost
Indirect Cost
Name
University of Southern California
Department
Type
Schools of Medicine
DUNS #
041544081
City
Los Angeles
State
CA
Country
United States
Zip Code
90089
Chen, M; Hieng, S; Qian, X et al. (1994) Regulation of hepatitis B virus ENI enhancer activity by hepatocyte-enriched transcription factor HNF3. Virology 205:127-32
Guo, W; Chen, M; Yen, T S et al. (1993) Hepatocyte-specific expression of the hepatitis B virus core promoter depends on both positive and negative regulation. Mol Cell Biol 13:443-8
Yeh, C T; Wong, S W; Fung, Y K et al. (1993) Cell cycle regulation of nuclear localization of hepatitis B virus core protein. Proc Natl Acad Sci U S A 90:6459-63
Liu, E S; Ou, J H; Lee, A S (1992) Brefeldin A as a regulator of grp78 gene expression in mammalian cells. J Biol Chem 267:7128-33
Xia, Y P; Yeh, C T; Ou, J H et al. (1992) Characterization of nuclear targeting signal of hepatitis delta antigen: nuclear transport as a protein complex. J Virol 66:914-21
Guo, W T; Bell, K D; Ou, J H (1991) Characterization of the hepatitis B virus EnhI enhancer and X promoter complex. J Virol 65:6686-92
Wang, J; Lee, A S; Ou, J H (1991) Proteolytic conversion of hepatitis B virus e antigen precursor to end product occurs in a postendoplasmic reticulum compartment. J Virol 65:5080-3
Guo, W T; Wang, J; Tam, G et al. (1991) Leaky transcription termination produces larger and smaller than genome size hepatitis B virus X gene transcripts. Virology 181:630-6
Yeh, C T; Ou, J H (1991) Phosphorylation of hepatitis B virus precore and core proteins. J Virol 65:2327-31
Ou, J H; Bao, H; Shih, C et al. (1990) Preferred translation of human hepatitis B virus polymerase from core protein- but not from precore protein-specific transcript. J Virol 64:4578-81

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