Troelsen Jesper T, Mitchelmore Cathy, Olsen Jørgen
Department of Medical Biochemistry and Genetics, Biochemical Laboratory C, The Panum Institute, University of Copenhagen, Blegdamsvej 3, DK-2200 N Copenhagen, Denmark.
Gene. 2003 Feb 13;305(1):101-11. doi: 10.1016/s0378-1119(02)01232-5.
Lactase phlorizin hydrolase is a small intestinal-specific brush border protein commonly used as a specific marker of differentiated enterocytes. A number of transcription factors involved in the enterocyte-specific expression of lactase phlorizin hydrolase have been identified. An upstream regulatory region, which we have named the "LPH enhancer", located at position -894 to -798 in the porcine lactase phlorizin hydrolase gene, is necessary for high differentiation-dependent LPH expression in intestinal cells. The LPH enhancer was studied by mutation analysis, transfection experiments and electrophoretical mobility shift assays. The LPH enhancer is active in intestinal cells (Caco-2) and not in non-intestinal cells (HeLa). The LPH enhancer is only able to enhance expression when it is located in front of an intestinal-specific promoter such as the lactase phlorizin hydrolase promoter or the sucrase-isomaltase promoter. In front of an SV40-derived promoter the LPH enhancer has no stimulatory effect. In addition to the lack of promoter-promiscuity, the LPH enhancer is not a classical enhancer in the sense that it is not orientation-independent and it cannot function when located 3' of a reporter gene. The LPH enhancer contains at least three cis-elements (at -894 to -880, -880 to -875 and -833 to -814) with functional importance for the LPH enhancer activity.
乳糖酶根皮苷水解酶是一种小肠特异性刷状缘蛋白,通常用作分化肠上皮细胞的特异性标志物。已经鉴定出许多参与乳糖酶根皮苷水解酶肠上皮细胞特异性表达的转录因子。位于猪乳糖酶根皮苷水解酶基因中-894至-798位置的上游调控区域,我们将其命名为“LPH增强子”,对于肠道细胞中高度分化依赖性LPH表达是必需的。通过突变分析、转染实验和电泳迁移率变动分析对LPH增强子进行了研究。LPH增强子在肠道细胞(Caco-2)中具有活性,而在非肠道细胞(HeLa)中无活性。LPH增强子只有位于肠道特异性启动子(如乳糖酶根皮苷水解酶启动子或蔗糖酶-异麦芽糖酶启动子)之前时才能增强表达。在SV40衍生的启动子之前,LPH增强子没有刺激作用。除了缺乏启动子通用性外,LPH增强子不是经典的增强子,因为它不是方向独立的,并且当位于报告基因的3'端时不能发挥作用。LPH增强子包含至少三个对LPH增强子活性具有功能重要性的顺式元件(位于-894至-880、-880至-875和-833至-814)。