GATA family transcription factors activate lactase gene promoter in intestinal Caco-2 cells
GATA family transcription factors activate lactase gene promoter in intestinal Caco-2 cells
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DOI：
10.1152/ajpgi.2001.280.1.g58
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发表时间：
2001-01-01
影响因子：
4.5
通讯作者：
Sibley, E
中科院分区：
文献类型：
--
作者：
Fang, RX;Olds, LC;Sibley, E
GATA转录因子家族在发育过程中调节特定组织的基因表达模式。我们研究了GATA蛋白与乳糖酶基因启动子之间的相互作用。结合乳糖酶基因GATA区顺式元件(-97~-73)的核蛋白用凝胶迁移率改变分析(EMSA)和GATA抗体超转移分析。用野生型和突变型荧光素酶启动子报告载体和GATA-4/5/6表达载体分别转染Caco-2细胞，检测乳糖酶启动子活性。EMSA与GATA区域探针产生需要GATA因子结合位点WGATAR的特定DNA-蛋白质复合体。该复合体被GATA-4和GATA-6特异性抗体识别。在Caco-2和非肠道QT6细胞中，GATA-4/5/6表达载体能够激活由野生型启动子驱动的转录，但不能激活由GATA结合位点突变的启动子驱动的转录。与乳糖酶顺式元件结合的GATA因子与功能启动子的激活相关。我们认为，在肠道中表达的GATA家族锌指蛋白GATA-4、-5和-6都可以与乳糖酶启动子GATA元件相互作用，并在Caco-2细胞中激活启动子。
The GATA family of transcription factors regulate tissue-specific patterns of gene expression during development. We have characterized the interaction between GATA proteins and the lactase gene promoter. Nuclear protein bound to the lactase gene GATA region cis element (-97 to -73) was analyzed by electrophoretic mobility shift assays (EMSA) and supershift assays with GATA antibodies. Lactase promoter activities were assayed in Caco-2 cells transfected with wild-type and mutated luciferase promoter-reporter constructs and GATA-4/ 5/6 expression constructs. EMSA with the GATA region probe yields a specific DNA-protein complex that requires the GATA factor binding site WGATAR. The complex is recognized by GATA-4- and GATA-6- specific antibodies. GATA-4/ 5/6 expression constructs are able to activate transcription driven by the wild-type promoter, but not by a promoter in which the GATA binding site is mutated, in Caco-2 and nonintestinal QT6 cells. GATA factor binding to the lactase cis element correlates with functional promoter activation. We conclude that each of the GATA family zinc finger proteins expressed in the intestine, GATA-4, -5, and -6, can interact with the lactase promoter GATA element and can function to activate the promoter in Caco-2 cells.