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Transcription factor Sp1 plays an important role in the regulation of copper homeostasis in mammalian cells.

机译:转录因子Sp1在调节哺乳动物细胞内铜稳态中起着重要作用。

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Copper is an essential metal nutrient, yet copper overload is toxic. Here, we report that human copper transporter (hCtr) 1 plays an important role in the maintenance of copper homeostasis by demonstrating that expression of hCtr1 mRNA was up-regulated under copper-depleted conditions and down-regulated under copper-replete conditions. Overexpression of full-length hCtr1 by transfection with a recombinant hCtr1 cDNA clone reduced endogenous hCtr1 mRNA levels, whereas overexpression of N terminus-deleted hCtr1 did not change endogenous hCtr1 mRNA levels, suggesting that increased functional hCtr1 transporter, which leads to increased intracellular copper content, down-regulates the endogenous hCtr1 mRNA. A luciferase assay using reporter constructs containing the hCtr1 promoter sequences revealed that three Sp1 binding sites are involved in the basal and copper concentration-dependent regulation of hCtr1 expression. Modulation of Sp1 levels affected the expression of hCtr1. We further demonstrated that the zinc-finger domain of Sp1 functions as a sensor of copper that regulates hCtr1 up and down in response to copper concentration variations. Our results demonstrate that mammalian copper homeostasis is maintained at the hCtr1 mRNA level, which is regulated by the Sp1 transcription factor.
机译:铜是必不可少的金属营养素,但铜超载却有毒。在这里,我们报告人类铜转运蛋白(hCtr)1在维持铜稳态中起着重要作用,这证明了hCtr1 mRNA的表达在缺铜条件下上调而在缺铜条件下下调。通过重组hCtr1 cDNA克隆转染来全长表达hCtr1会降低内源性hCtr1 mRNA水平,而过表达N末端缺失的hCtr1不会改变内源性hCtr1 mRNA水平,这表明功能性hCtr1转运蛋白增加,导致细胞内铜含量增加,下调内源性hCtr1 mRNA。使用包含hCtr1启动子序列的报告基因构建体进行的荧光素酶分析显示,三个Sp1结合位点参与了hCtr1表达的基础和铜浓度依赖性调节。 Sp1水平的调节影响hCtr1的表达。我们进一步证明了Sp1的锌指结构域充当铜的传感器,该铜可响应铜浓度的变化而上下调节hCtr1。我们的结果表明,哺乳动物铜稳态保持在hCtr1 mRNA水平,该水平由Sp1转录因子调控。

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