Short J M, Wynshaw-Boris A, Short H P, Hanson R W
J Biol Chem. 1986 Jul 25;261(21):9721-6.
Hormonal regulatory elements within the phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.1.32) (PEPCK) promoter region were mapped using a series of 5' deletions linked to the amino-3'-glycosyl phosphotransferase structural gene. These deletion mutants were stably transfected into the genome of FTO-2B hepatoma cells. A 47-base pair region of the PEPCK promoter was identified which was essential for stimulation by dibutyryl cAMP. A 12-base pair core sequence (CTTACGTCAGAG) within this region shows significant homology with sequences in four other cAMP-regulated genes. There are two glucocorticoid regulatory elements within the promoter, as well as an inhibitory element which depresses the level of basal gene transcription. The deletion of this inhibitory sequence prevents the induction of the chimeric gene by dexamethasone. The existence of the hormone regulatory domains within the PEPCK promoter was confirmed by attaching these elements upstream of the heterologous Herpes simplex virus thymidine kinase structural gene, containing its own promoter.
利用一系列与氨基-3'-糖基磷酸转移酶结构基因相连的5'端缺失片段，对磷酸烯醇丙酮酸羧激酶（GTP）（EC 4.1.1.32）（PEPCK）启动子区域内的激素调控元件进行了定位。这些缺失突变体被稳定转染到FTO-2B肝癌细胞基因组中。确定了PEPCK启动子的一个47个碱基对的区域，该区域对于二丁酰环磷腺苷（dbcAMP）的刺激至关重要。该区域内一个12个碱基对的核心序列（CTTACGTCAGAG）与其他四个环磷腺苷调节基因的序列具有显著同源性。启动子内有两个糖皮质激素调控元件，以及一个抑制元件，该抑制元件可降低基础基因转录水平。该抑制序列的缺失可阻止地塞米松对嵌合基因的诱导。通过将这些元件连接到含有自身启动子的异源单纯疱疹病毒胸苷激酶结构基因的上游，证实了PEPCK启动子内激素调控结构域的存在。
