Genomic structure and promoter activity of the mouse polysialic acid synthase gene (mST8Sia II) . Brain-specific expression from a TATA-less GC-rich sequence

Yoshida, Y.; Kurosawa, N.; Kanematsu, T.; Kojima, N.; Tsuji, S.

Journal of Biological Chemistry 271(47): 30167-30173

1996


ISSN/ISBN: 0021-9258
PMID: 8939967
DOI: 10.1074/jbc.271.47.30167
Accession: 002852089

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Abstract
The mouse ST8Sia II (mST8Sia II/STX) gene encodes a neural cell adhesion molecule-specific polysialic acid synthase whose expression is regulated during the developmental stages of mouse brain. Tissue-specific and developmentally-regulated gene expression were investigated by isolating the complete genomic DNA and characterizing the promoter of the gene for mST8Sia II. The mST8Sia II gene is ~80 kb and composed of 6 exons (EMBL/GenBank accession number X83562 and X99645-X99651). Primer extension and S1 nuclease protection analyses revealed that transcription started 167 nucleotides upstream of the translation initiation site. Promoter analysis of the 5'-flanking region of the mST8Sia II gene using a luciferase gene reporter system revealed strong promoter activity in retinoic acid-induced differentiated P19 cells, which highly express the mST8Sia II gene. Deletion analysis demonstrated that the minimal promoter activity detected for the proximal region 325 bp upstream from the translational initiation codon (-158 to +167) could be modulated by various sequences within the 9.5-kb 5'-flanking region. The minimal promoter was embedded in a GC-rich domain (74%, GC content), in which 2 Sp1 binding motifs as well as a long purine-rich region were found, but it lacked TATA and CAAT boxes. The positive regulatory region located between -159 and -659 contained 2 additional Sp1 binding motifs and a long pyrimidine-rich region. The minimal promoter region of the mST8Sia II gene was found to be sufficient to express a reporter gene in mST8Sia II gene-expressing neural differentiated P19 cells but not in non-expressing ones. Thus, the TATA-less GC-rich minimal promoter region of mST8Sia II probably controls the cell type-specific expression of the gene.