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Cloning of the mouse somatostatin receptor subtype 5 gene: Promoter structure and function

Lookup NU author(s): Dr Robert James


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Somatostatin is a peptide hormone whose actions are mediated by five somatostatin receptor subtypes (sst1-5). In the pituitary, somatostatin inhibits TSH release from thyrotropes and GH release from somatotropes. We have shown that sst5 transcripts and protein are induced by thyroid hormone in TtT-97 thyrotropic tumors. To map sequences responsible for promoter activity in pituitary cells, we cloned the mouse sst5 coding region of 362 amino acids and 12 kb of upstream DNA. Initial transfection studies in TtT-97 or GH3 cells mapped high levels of basal promoter activity to a 5.6-kb fragment upstream of the translational start, whereas shorter genomic fragments had low activity. To identify the transcriptional start site we used 5' RACE with TtT-97 poly A+RNA and a sst5 antisense coding region primer. Sequence comparison between the complementary DNA and the gene revealed that the mouse sst5 gene contains 3 exons and 2 introns. The entire coding region was contained in exon 3. Two differently sized RACE products demonstrated alternate exon splicing of two untranslated exons in TtT-97 cells. A promoter fragment from -290/+48 linked to a luciferase reporter demonstrated 600- and 900-fold higher activity over a promoterless control in GH3 mammosomatotropes and TtT-97 thyrotropes, respectively, whereas a larger fragment extending to -6400 exhibited no additional promoter activity. Cloning of the sst5 gene will facilitate the mapping of basal and regulated responses at the transcriptional level.

Publication metadata

Author(s): James RA; Gordon DF; Woodmansee WW; Lewis SR; Wood WM; Ridgway EC

Publication type: Article

Publication status: Published

Journal: Endocrinology

Year: 1999

Volume: 140

Issue: 12

Pages: 5598-5608

Print publication date: 01/12/1999

ISSN (print): 0013-7227

ISSN (electronic): 1945-7170

Publisher: The Endocrine Society


DOI: 10.1210/en.140.12.5598

PubMed id: 10579323


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