Characterization of the 5′‐flanking promoter region of the rat somatostatin receptor subtype 3 gene

FEBS Letters

Glos, Michael¹  Kreienkamp, Hans-Jürgen¹  Hausmann, Holger¹  Richter, Dietmar¹ 

[1] Institut für Zellbiochemie und klinische Neurobiologie, UKE, Universität Hamburg, Martinistraße 52, 20246 Hamburg, Germany

关键词: Somatostatin;    Promoter;    5′ Untranslated region;    5′ Rapid amplification of cDNA ends;    HEK;    human embryonic kidney;    NRSE;    neuronal restrictive silencing element;    RACE;    rapid amplification of cDNA ends;    SST;    somatostatin;    SSTR;    somatostatin receptor;    UTR;    untranslated region;   

DOI  :  10.1016/S0014-5793(98)01411-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We investigated the 5′-flanking promoter region of the rat somatostatin receptor subtype 3 (rSSTR3). Using a cDNA probe, genomic clones containing the 5′-flanking promoter region of the rSSTR3 gene were isolated. A sequence of 5.4 kb directly upstream from the start codon was analyzed and two introns were found in the 5′ untranslated region (UTR) of the cDNA sequence. The transcriptional initiation site was determined by 5′ rapid amplification of cDNA ends (RACE), primer extension and RNase protection analysis with cerebellar RNA. Two major transcriptional initiation sites were found at position −1040 (tsp1) and −856 (tsp2) relative to the translational initiation site. Like a number of other promoters of G-protein-coupled receptors, the rSSTR3 gene lacks TATA and CAAT motifs and includes G+C-rich regions. Functional analysis of the promoter region by transfecting rSSTR3 luciferase-reporter gene constructs into rat pituitary GH₃ cells and HEK 293 cells indicated that a 107-bp region upstream of tsp2 was sufficient to drive transcription. Furthermore a 562-bp region at position −1304 to −1865 upstream of the ATG start codon exerted a negative regulatory effect on transcriptional activity.

【 授权许可】

Unknown