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Cloning and characterization of the human beta2-glycoprotein I (beta2-GPI) gene promoter: roles of the atypical TATA box and hepatic nuclear factor-1alpha in regulating beta2-GPI promoter activity.
Biochem J. 2004 Jun 01; 380(Pt 2):455-63.BJ

Abstract

Beta2-glycoprotein I (beta2-GPI) is a plasma glycoprotein primarily synthesized in the liver. The interindividual variability of beta2-GPI expression in subjects with various metabolic syndromes and disease states suggests that it may have clinical importance. However, the regulation of beta2-GPI gene expression has not been clarified. To gain more insight into the control of beta2-GPI gene expression, we cloned the 4.1-kb 5'-flanking region and characterized the proximal promoter of the beta2- GPI gene in this study. Cis -acting elements required for beta2-GPI promoter activity were identified with transient transfection assays in the hepatoma cell lines HepG2 and Huh7 and in non-hepatic HeLa cells. Serial deletion analyses of the beta2-GPI 5'-flanking sequence revealed that the region from -197 to +7 had strong promoter activity in hepatoma cells but not in HeLa cells. Truncation and site-directed mutagenesis of putative cis -elements within this region showing an atypical TATA box and a HNF-1 (hepatic nuclear factor-1) element were both essential for the beta2-GPI promoter activity. Subsequent gel mobility shift assays confirmed the interaction of HNF-1alpha with the HNF-1 site residing downstream of the TATA box. Co-transfection of beta2-GPI promoter-luciferase vector with HNF-1alpha expression vector in Huh7 and HNF-1-deficient HeLa cells demonstrated the transactivation effect of HNF-1alpha on beta2-GPI promoter activity. In addition, overexpression of HNF-1alpha enhanced the endogenous beta2-GPI expression. These results suggest that the atypical TATA box and HNF-1 cis-element are critical for beta2-GPI transcription and HNF-1alpha may play an important role in cell-specific regulation of beta2-GPI gene expression.

Authors+Show Affiliations

Institute of Biochemistry, School of Life Science, National Yang-Ming University, Taipei 112, Taiwan, ROC.No affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

14984368

Citation

Wang, Hsueh-Hsiao, and An-Na Chiang. "Cloning and Characterization of the Human Beta2-glycoprotein I (beta2-GPI) Gene Promoter: Roles of the Atypical TATA Box and Hepatic Nuclear Factor-1alpha in Regulating beta2-GPI Promoter Activity." The Biochemical Journal, vol. 380, no. Pt 2, 2004, pp. 455-63.
Wang HH, Chiang AN. Cloning and characterization of the human beta2-glycoprotein I (beta2-GPI) gene promoter: roles of the atypical TATA box and hepatic nuclear factor-1alpha in regulating beta2-GPI promoter activity. Biochem J. 2004;380(Pt 2):455-63.
Wang, H. H., & Chiang, A. N. (2004). Cloning and characterization of the human beta2-glycoprotein I (beta2-GPI) gene promoter: roles of the atypical TATA box and hepatic nuclear factor-1alpha in regulating beta2-GPI promoter activity. The Biochemical Journal, 380(Pt 2), 455-63.
Wang HH, Chiang AN. Cloning and Characterization of the Human Beta2-glycoprotein I (beta2-GPI) Gene Promoter: Roles of the Atypical TATA Box and Hepatic Nuclear Factor-1alpha in Regulating beta2-GPI Promoter Activity. Biochem J. 2004 Jun 1;380(Pt 2):455-63. PubMed PMID: 14984368.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Cloning and characterization of the human beta2-glycoprotein I (beta2-GPI) gene promoter: roles of the atypical TATA box and hepatic nuclear factor-1alpha in regulating beta2-GPI promoter activity. AU - Wang,Hsueh-Hsiao, AU - Chiang,An-Na, PY - 2004/02/25/accepted PY - 2004/02/19/revised PY - 2003/10/22/received PY - 2004/2/27/pubmed PY - 2004/10/19/medline PY - 2004/2/27/entrez SP - 455 EP - 63 JF - The Biochemical journal JO - Biochem J VL - 380 IS - Pt 2 N2 - Beta2-glycoprotein I (beta2-GPI) is a plasma glycoprotein primarily synthesized in the liver. The interindividual variability of beta2-GPI expression in subjects with various metabolic syndromes and disease states suggests that it may have clinical importance. However, the regulation of beta2-GPI gene expression has not been clarified. To gain more insight into the control of beta2-GPI gene expression, we cloned the 4.1-kb 5'-flanking region and characterized the proximal promoter of the beta2- GPI gene in this study. Cis -acting elements required for beta2-GPI promoter activity were identified with transient transfection assays in the hepatoma cell lines HepG2 and Huh7 and in non-hepatic HeLa cells. Serial deletion analyses of the beta2-GPI 5'-flanking sequence revealed that the region from -197 to +7 had strong promoter activity in hepatoma cells but not in HeLa cells. Truncation and site-directed mutagenesis of putative cis -elements within this region showing an atypical TATA box and a HNF-1 (hepatic nuclear factor-1) element were both essential for the beta2-GPI promoter activity. Subsequent gel mobility shift assays confirmed the interaction of HNF-1alpha with the HNF-1 site residing downstream of the TATA box. Co-transfection of beta2-GPI promoter-luciferase vector with HNF-1alpha expression vector in Huh7 and HNF-1-deficient HeLa cells demonstrated the transactivation effect of HNF-1alpha on beta2-GPI promoter activity. In addition, overexpression of HNF-1alpha enhanced the endogenous beta2-GPI expression. These results suggest that the atypical TATA box and HNF-1 cis-element are critical for beta2-GPI transcription and HNF-1alpha may play an important role in cell-specific regulation of beta2-GPI gene expression. SN - 1470-8728 UR - https://www.unboundmedicine.com/medline/citation/14984368/Cloning_and_characterization_of_the_human_beta2_glycoprotein_I__beta2_GPI__gene_promoter:_roles_of_the_atypical_TATA_box_and_hepatic_nuclear_factor_1alpha_in_regulating_beta2_GPI_promoter_activity_ DB - PRIME DP - Unbound Medicine ER -