title: The hepatitis B virus core promoter is strongly activated by the liver nuclear receptor fetoprotein transcription factor or by ectopically expressed steroidogenic factor 1.
abstract: Orphan nuclear receptor fetoprotein transcription factor (FTF) was previously identified as a specific regulator of the alpha(1)-fetoprotein gene during early liver development and in response to hormonal signals (L. Galarneau, J.-F. Paré, D. Allard, D. Hamel, L. Lévesque, J. D. Tugwood, S. Green, and L. Bélanger, Mol. Cell. Biol. 16:3853-3865, 1996). Here we report a functional analysis of FTF interactions with the hepatitis B virus (HBV) nucleocapsid promoter. DNA-protein-binding assays show that the HBV core promoter contains two high-affinity FTF-binding sites and a third, lower-affinity site shared with other receptors. Transfections in HepG2, Hep3B, and PLC/PRF/5 hepatoma cells using chloramphenicol acetyltransferase reporter genes with the nucleocapsid promoter linked or not linked to enhancer I indicate that FTF is a potent activator of the HBV core promoter, more efficient than HNF4alpha, HNF3alpha, HNF3beta, or C/EBPalpha. Steroidogenic factor 1, a close FTF homolog which binds to the same DNA motif and is expressed ectopically in HepG2 cells, seems to be an even stronger inducer than FTF. Point mutations of the FTF-binding sites indicate direct FTF activatory effects on the core promoter and the use of both high-affinity sites for productive interaction between the core promoter and enhancer I. Coexpression assays further indicate that FTF and HNF4alpha are the most efficient partners for coactivation of the pregenomic core promoter, which may largely account for the hepatic tropism and the early amplification of HBV infection. Carboxy terminus-truncated FTF behaves as a dominant negative mutant to compete all three FTF sites and strongly deactivate core promoter interactions with enhancer I; this suggests possible new ways to interfere with HBV infection.
authors.affiliation: Le Centre de Recherche en Cancérologie de l'Université Laval, L'Hôtel-Dieu de Québec, Département de Biologie Médicale, Faculté de Médecine, Québec G1R 2J6, Canada.
authors.forename: S
authors.initials: S
authors.lastname: Gilbert
authors.forename: L
authors.initials: L
authors.lastname: Galarneau
authors.forename: A
authors.initials: A
authors.lastname: Lamontagne
authors.forename: S
authors.initials: S
authors.lastname: Roy
authors.forename: L
authors.initials: L
authors.lastname: Bélanger
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D004279:DNA, Viral; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004742:Enhancer Elements, Genetic; D051717:Fushi Tarazu Transcription Factors; D006515:Hepatitis B virus / Q000235:genetics*; D018398:Homeodomain Proteins; D006801:Humans; D008099:Liver / Q000378:metabolism; D008969:Molecular Sequence Data; D019251:Nucleocapsid / Q000235:genetics*; D019590:Nucleocapsid Proteins; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D054339:Steroidogenic Factor 1; D014157:Transcription Factors / Q000378:metabolism*; D015533:Transcriptional Activation; D014407:Tumor Cells, Cultured; D000509:alpha-Fetoproteins
chemical_list: D004279:DNA, Viral; D004268:DNA-Binding Proteins; D051717:Fushi Tarazu Transcription Factors; D018398:Homeodomain Proteins; C515057:NR5A1 protein, human; D019590:Nucleocapsid Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D054339:Steroidogenic Factor 1; D014157:Transcription Factors; D000509:alpha-Fetoproteins; C099948:fetoprotein transcription factor; C099600:nucleocapsid protein, Hepatitis virus