Raychowdhury Raktima, Fleming John V, McLaughlin John T, Bulitta Clemens J, Wang Timothy C
Harvard Medical School and Gastrointestinal Unit, Massachusetts General Hospital, Boston, MA 02114, USA.
Biochem Biophys Res Commun. 2002 Oct 11;297(5):1089-95. doi: 10.1016/s0006-291x(02)02345-8.
In human gastric cancer cells the human histidine decarboxylase gene is regulated by gastrin through two overlapping cis-acting elements known as gastrin response elements 1&2 (GAS-RE1, GAS-RE2) [J. Biol. Chem. 274 (1999) 20961]. Here, we report the identification and characterization of a third element GAS-RE3 that was localized to a region +28 to +48 downstream of the transcriptional start site (+1). Gastrin stimulation induced a rapid increase in binding to the element of a novel nuclear factor named gastrin response element-binding protein 3 (GAS-REBP3). Block mutations in the GAS-RE3 sequence (+38GTGCG(+42) to +38TAAGT(+42)) led to reduced promoter activity and decreased binding in EMSA. UV cross-linking studies and Southwestern blot analysis with wildtype and mutant GAS-RE3 showed that GAS-REBP3 was a approximately 110kDa protein. Thus, gastrin-mediated regulation of HDC gene expression appears to be mediated by a complex cis-acting element, which binds at least three distinct nuclear factors.
在人胃癌细胞中,人组氨酸脱羧酶基因受胃泌素通过两个重叠的顺式作用元件调控,这两个元件被称为胃泌素反应元件1和2(GAS - RE1、GAS - RE2)[《生物化学杂志》274 (1999) 20961]。在此,我们报告了第三个元件GAS - RE3的鉴定与特性,它定位于转录起始位点(+1)下游+28至+48区域。胃泌素刺激导致一种名为胃泌素反应元件结合蛋白3(GAS - REBP3)的新型核因子与该元件的结合迅速增加。GAS - RE3序列(+38GTGCG(+42)至+38TAAGT(+42))中的阻断突变导致启动子活性降低以及电泳迁移率变动分析(EMSA)中的结合减少。对野生型和突变型GAS - RE3进行的紫外线交联研究和蛋白质印迹分析表明,GAS - REBP3是一种约110kDa的蛋白质。因此,胃泌素介导的HDC基因表达调控似乎是由一个复杂的顺式作用元件介导的,该元件可结合至少三种不同的核因子。