Transcription factor Sp1 activates the expression of the mouse tenascin-X gene.
Tenascin-X (TNX) is an extracellular matrix protein that is prominent in the heart and muscle. We previously showed that TNX is expressed in fibroblast cells in culture. To elucidate the molecular basis of the TNX gene expression, the promoter region of the mouse TNX gene (mTnx) has been characterized. The two adjacent transcription initiation sites were identified at 68 and 67 bp upstream of the previously known 5'-untranslated exon. Transient transfection of L and 293T cells with 5'-deletion constructs of the promoter region linked to the luciferase reporter revealed that the region (-141 to -136) containing a transcription factor Sp1-binding element contributes to the expression of mTnx. Site-directed mutagenesis of the Sp1-binding region confirmed this result. Electrophoretic mobility shift analysis using nuclear extracts obtained from the cells demonstrated that a distinct Sp1-DNA complex is formed at the element. Our results show that Sp1 plays a critical role in the gene expression of mTnx.
Minamitani
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Department of Molecular Biology, Hokkaido University, Sapporo, 060-0812, Japan.
Ariga
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Matsumoto
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Ki
D000818:Animals; D001482:Base Composition; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006801:Humans; D007739:L Cells; D008156:Luciferases / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D019063:Tenascin / Q000235:genetics*; D014162:Transfection
D017931:DNA Primers; D016329:Sp1 Transcription Factor; D019063:Tenascin; C081865:tenascin X; D004247:DNA; D008156:Luciferases