Differential allelic expression of dopamine D1 receptor gene (DRD1) is modulated by microRNA miR-504.
Differential allelic expression of dopamine D1 receptor gene (DRD1) is modulated by microRNA miR-504.
复制标题
DOI：
10.1016/j.biopsych.2008.11.024
复制
发表时间：
2009-04-15
影响因子：
10.6
通讯作者：
Li, Ming D.
中科院分区：
文献类型：
--
作者：
Huang, Weihua;Li, Ming D.
关键词：
以前，我们不仅报道了多巴胺D_1受体基因(DRD_1)与尼古丁依赖(ND)相关，而且发现DRD_1的3‘-非翻译区(3’-UTR)上rs686的两个等位基因(A和G)存在差异表达。然而，这种差异表达的机制仍有待确定。我们假设它是由miRNA靶向引起的。我们首先使用MicroInspector算法来识别可能针对DRD13‘UTR中rs686多态的microRNAs(MiRNAs)，然后使用荧光素酶报告基因分析结合定点突变来测试所预测的miRNA靶向性。我们还通过基因表达实验检测了DRD1的miRNA靶向性。在通过计算分析预测的两个miRNAs中，我们发现miR-504而不是miR-296通过靶向DRD13‘UTR上调报告荧光素酶的活性和增加DRD1的表达，而抑制miR-504而不是miR-296具有相反的作用。此外，我们还发现，定点突变证实miR-504与DRD13‘非编码区的直接结合导致了两个等位基因之间的显著表达差异。MIR-504通过直接与3‘端非编码区结合上调DRD1的表达，从而导致DRD1的差异等位基因特异性表达。
Previously, we not only reported that dopamine D1 receptor gene (DRD1) is associated with nicotine dependence (ND), but demonstrated that two alleles (A and G) of polymorphism rs686 in the 3′-untranslated region (3′UTR) of DRD1 are expressed differentially. However, the mechanism underlying the differential expression remains to be determined. We hypothesize that it is caused by miRNA targeting. We first used the MicroInspector algorithm to identify microRNAs (miRNAs) potentially targeting the rs686 polymorphism in the DRD1 3′UTR, and then employed a luciferase reporter assay combined with site-directed mutagenesis to test the predicted miRNA targeting. We also examined the miRNA targeting of DRD1 with a gene expression assay. Of two miRNAs predicted by computational analyses, we found that miR-504, not miR-296, up-regulated reporter luciferase activity and increased DRD1 expression by targeting the DRD1 3′UTR, whereas inhibition of miR-504, not miR-296, had the opposite effect. Furthermore, we revealed that the direct binding of miR-504 to the DRD1 3′UTR, verified by site-directed mutagenesis, caused a significant expression difference between the two alleles. miR-504 up-regulates DRD1 expression by direct binding to the 3′UTR, which leads to differential allele-specific expression of DRD1.
登录
查看更多内容
影响因子：
14.9
作者：
Rusinov, V;Baev, V;Minkov, IN;Tabler, M
通讯作者：
Tabler, M
影响因子：
56.9
作者：
Vasudevan, Shobha;Tong, Yingchun;Steitz, Joan A.
通讯作者：
Steitz, Joan A.
影响因子：
5.3
作者：
Huang, Weihua;Ma, Jennie Z.;Li, Ming D.
通讯作者：
Li, Ming D.
影响因子：
56.9
作者：
Frayling, Timothy M.;Timpson, Nicholas J.;McCarthy, Mark I.
通讯作者：
McCarthy, Mark I.
影响因子：
5.3
作者：
Gecz, J;Baker, E;Mulley, JC
通讯作者：
Mulley, JC