Regulation of the rat S100 beta gene expression: the role of the 2 kb 5'-upstream sequence in glial specific expression.
We have studied the role of the 2047 bp 5'-upstream region and 232 bp first exon sequence of the rat S100 beta gene in glial specific expression. S100 beta luciferase expression vectors carrying serial deletions of the S100 beta 5' upstream sequence were constructed and transiently transfected into the peripheral nervous system (PNS) glial-type cell line RT4-D6, the PNS neuronal-type cell line RT4-E5, and the central nervous system (CNS) glial-type cell line C6. The hepatoma cell line HTC was also transfected as a nonneural control. From this functional analysis, we found a glial-specific positive regulatory sequence(s) mapped between -583 and -106 relative to the transcriptional start site. This region confers a significantly higher level of luciferase expression in the glial-type cell lines RT4-D6 and C6 than in the neuronal cell line RT4-E5 and the hepatoma cell line HTC. Also, a non-cell type specific positive regulatory element was identified in the first exon sequence between +78 and +232. Though non-cell type-specific, it was found to have a predominant effect in glial cells. From these observations, we have concluded that the 2047 bp 5'-upstream region and 232 bp of the first exon sequence confers the high levels of S100 beta expression in glial cells through these two positive elements.
Hagiwara
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Department of Molecular, Cellular and Developmental Biology, University of Colorado at Boulder 80309, USA.
Sueoka
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D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D017931:DNA Primers; D005091:Exons / Q000235:genetics; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D008156:Luciferases / Q000096:biosynthesis; D008969:Molecular Sequence Data; D009457:Neuroglia / Q000378:metabolism*; D051381:Rats; D012045:Regulatory Sequences, Nucleic Acid; D009418:S100 Proteins / Q000096:biosynthesis / Q000235:genetics*; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D009418:S100 Proteins; D004247:DNA; D008156:Luciferases