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首页> 外文期刊>American Journal of Physiology >GATA family transcription factors activate lactase gene promoter in intestinal Caco-2 cells.
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GATA family transcription factors activate lactase gene promoter in intestinal Caco-2 cells.

机译：GATA家族转录因子激活肠道Caco-2细胞中的乳糖酶基因启动子。

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摘要

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.
机译：GATA转录因子家族在发育过程中调节基因表达的组织特异性模式。我们已经表征了GATA蛋白和乳糖酶基因启动子之间的相互作用。结合乳糖酶基因GATA区域顺式元件（-97至-73）的核蛋白通过电泳迁移率变动分析（EMSA）和GATA抗体的超转变分析进行了分析。在用野生型和突变的荧光素酶启动子-报告子构建体和GATA-4 / 5/6表达构建体转染的Caco-2细胞中测定了乳糖酶启动子活性。带有GATA区域探针的EMSA产生需要GATA因子结合位点WGATAR的特定DNA-蛋白质复合物。该复合物可被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表达构建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因子与功能性启动子激活相关。我们得出的结论是，在肠中表达的每个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，并可以激活Caco-2细胞中的启动子。

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