TAILIEUCHUNG - Báo cáo khoa học: Characterization of the promoter for the mouse a3 integrin gene Involvement of the Ets-family of transcription factors in the promoter activity

Thea3b1 integrin is an adhesion receptor for extracellular matrix proteins including isoforms of laminin, and the changes of its expression level in various cancer cells are thought to cause their malignant phenotypes. We have cloned an approximately 4 kb DNA fragment of the 5¢-flanking region of the murinea3 integrin gene and analyzed its promoter activity. | Eur. J. Biochem. 269 4524-4532 2002 FEBS 2002 doi Characterization of the promoter for the mouse a3 integrin gene Involvement of the Ets-family of transcription factors in the promoter activity Takumi Kato1 Kouji Katabami1 Hironori Takatsuki1 Seon Ae Han2 Ken-ichi Takeuchi2 Tatsuro Irimura2 and Tsutomu Tsuji1 2 1 Department of Microbiology Hoshi University School of Pharmacy and Pharmaceutical Sciences Tokyo Japan 2Laboratory of Cancer Biology and Molecular Immunology Graduate School of Pharmaceutical Sciences University of Tokyo Japan The a3p1 integrin is an adhesion receptor for extracellular matrix proteins including isoforms of laminin and the changes of its expression level in various cancer cells are thought to cause their malignant phenotypes. We have cloned an approximately 4 kb DNA fragment of the 5 -flanking region of the murine a3 integrin gene and analyzed its promoter activity. Transfection of MKN1 gastric carcinoma cells with serially truncated segments of the 5 -flanking region linked to a luciferase gene indicated that a 537-bp SalI SacI fragment upstream of exon 1 was sufficient to promote high level gene expression. By 5 -rapid amplification of cDNA ends 5 -RACE using a cap site-labeled cDNA library we determined one major and one minor transcription start sites in this region. The murine a3 integrin gene was found to contain a CCAAT box but to lack a TATA box. Luciferase assay following transfection with a series of deletion constructs of the SalI SacI fragment revealed that the sequence between positions -260 and -119 bp relative to the major transcription start site is required for efficient transcription in gastric carcinoma cells. The sequence analysis of this segment showed the presence of several consensus sequences for transcription factors including Ets GATA and MyoD E-box binding factors. The introduction of mutation in one of the Ets-binding sequences greatly decreased its promoter activity suggesting .

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