Transcriptional regulation of the major zinc uptake protein hZip1 in prostate cancer cells.
Transcriptional regulation of the major zinc uptake protein hZip1 in prostate cancer cells.
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DOI：
10.1016/j.gene.2008.10.015
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发表时间：
2009-02-15
期刊：
影响因子：
3.5
通讯作者：
Kolenko, Vladimir M.
中科院分区：
文献类型：
--
作者：
Makhov, Peter;Golovine, Konstantin;Uzzo, Robert G.;Wuestefeld, Torsten;Scoll, Benjamin J.;Kolenko, Vladimir M.
hZip 1是调节前列腺细胞锌积累的主要锌摄取转运蛋白。调节hZip 1表达的机制尚未描述。为了探索hZip 1基因的转录调控机制，我们确定了假定的hZip 1启动子序列，并确定了预测的hZip 1启动子区域内的潜在转录起始位点。为了进一步表征基础hZip 1转录的启动子区域，通过PCR扩增产生3′和5′缺失构建体和具有推定转录因子的突变结合位点的构建体，并在PC-3前列腺癌细胞中用荧光素酶报告基因测定评估转录活性。特异性转录因子结合hZip 1核心启动子的能力通过EMSA、GelSupershift和ChIP测定来证实。我们的实验确定了负责hZip 1组成性表达的核心启动子区域，并证明了SP1和CREB 1在前列腺癌细胞中hZip 1基因转录调节中的关键作用。
hZip1 has been characterized as the major zinc uptake transporter regulating the accumulation of zinc in prostate cells. The mechanisms regulating expression of hZip1 have not been described. To explore the mechanisms of transcriptional regulation of the hZip1 gene, we determined the putative promoter sequence for hZip1 and identified the potential transcription start site within the predicted hZip1 promoter region. To further characterize the promoter region for basal hZip1 transcription, 3′ and 5′ deletion constructs and constructs with mutated binding sites for putative transcription factors were generated by PCR amplification and assessed for transcriptional activity with a luciferase reporter assay in PC-3 prostate cancer cells. The ability of the specific transcription factors to bind the hZip1 core promoter was confirmed by EMSA, Gel Supershift and ChIP assays. Our experiments identified the core promoter region responsible for constitutive expression of hZip1 and demonstrated critical roles for SP1 and CREB1 in transcriptional regulation of the hZip1 gene in prostate cancer cells.
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