| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1997: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1996: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Research Abstract |
Adrenocorticotropin receptor (ACTH-R) is a 7-transmembrane spanning protein through which ACTH signal is transmitted into the cell to mount steroidogenesis in the adrenocortical cells. We demonstrated recently that the mouse ACTH-R gene consists of four exons (Shimizu et al., Gene, 1997) : the first three exons encode 5'-untranslated region (UTR) and the fourth exon encodes part of 5'-UTR,the entire coding region, and the whole of 3'-UTR.Northern blot analysis demonstrated 1.8kb mRNA in mouse adrenal which consists of two mRNA species, one with and one without 57-bp exon 2 by alternative splicing.
Next, primer extension experiments were done and four putative transcription initiation sites spanning 124 bp were identified. When the initiation site most downstream was designated +1, three other sites were located at-62, -94, and-123, and exon 1 was of 50 bp almost the same length as that of exon 1 of the human ACTH-R gene. Genomic sequence analysis revealed that consensus sequence for TATA-, CAAT-, and GC-box was absent within 50 bp upstream of each of four putative transcriprion initiation sites. Functional promoter analysis of the mouse ACTH-R gene, using luciferase chimeric plasmids and Y-1 mouse adrenocortical cells revealed the minimal promoter to be located within-62 bp and tissue specific regulatory element (s) of the ACTH-R gene expression within-195 bp upstream of the most downstrewam transcription initiation site.
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