Membrane-initiated estradiol signaling increases tyrosine hydroxylase promoter activity with ER alpha in PC12 cells.
Membrane-initiated estradiol signaling increases tyrosine hydroxylase promoter activity with ER alpha in PC12 cells.
复制标题
PC12 细胞中膜启动的雌二醇信号通过 ER α 增加酪氨酸羟化酶启动子活性。
DOI：
10.1111/j.1471-4159.2009.06430.x
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发表时间：
2010
影响因子：
4.7
通讯作者：
Sabban,EstherL
中科院分区：
文献类型：
--
作者：
Maharjan,Shreekrishna;Serova,LidiaI;Sabban,EstherL
神经化学杂志。（2010）112，42- 55. AbstractTyrosine hydroxylase（TH）promoter activity is induced by 17β-estradiol（E2）in PC12 cells expressing estradiol receptor-alpha（ERα）requiring a cAMP/calcium response element（CRE/CaRE）at −45.为了检查膜启动的雌二醇信号传导是否是这种诱导的基础，将TH报告基因构建体和ERα表达载体共转染的细胞暴露于膜不透性雌二醇缀合物（β-雌二醇-6-（O-羧基-甲基）肟-牛血清白蛋白，E2 BSA）。E2 BSA可剂量依赖性和时间依赖性地提高TH启动子活性。E2 BSA还引起CRE结合蛋白（CREB）的快速磷酸化，并增加CRE驱动的启动子活性。CREB的显性阴性形式的过度表达，在DNA结合或磷酸化位点的突变，阻止TH启动子响应E2 BSA。用蛋白激酶A（PKA）和MEK抑制剂预处理减少了CREB和ERK的E2依赖性磷酸化，也减少了E2或E2 BSA对TH启动子活性的诱导。用C451 A突变阻断ERα的S-棕榈酰化和/或用2-溴棕榈酸酯预处理不能阻止但反而增强了E2或E2 BSA诱导的TH启动子活性。这些发现首次揭示，雌二醇诱导PC 12细胞中TH基因与ERα的转录涉及膜启动的雌二醇信号传导，快速激活PKA/MEK双重信号传导途径，导致CREB磷酸化，作用于CRE/CaRE。这些数据表明雌二醇影响体内儿茶酚胺能系统的可能机制。
J. Neurochem.(2010)112, 42–55.AbstractTyrosine hydroxylase (TH) promoter activity is induced by 17β‐estradiol (E2) in PC12 cells expressing estradiol receptor‐alpha (ERα) requiring a cAMP/calcium response element (CRE/CaRE) at −45. To examine whether membrane‐initiated estradiol signaling is underlying this induction, cells co‐transfected with TH reporter construct and ERα expression vector were exposed to membrane‐impermeant estradiol conjugate (β‐estradiol‐6‐(O‐carboxy‐methyl) oxime‐bovine serum albumin, E2BSA). TH promoter activity was elevated by E2BSA in dose‐ and time‐dependent manner. E2BSA also elicited rapid phosphorylation of CRE binding protein (CREB) and increased CRE‐driven promoter activity. Over‐expression of dominant negative forms of CREB, with mutations in DNA binding or phosphorylation site, prevented TH promoter response to E2BSA. Pre‐treatment with protein kinase A (PKA) and MEK inhibitors reduced E2dependent phosphorylation of CREB and ERK, and also decreased induction of TH promoter activity by E2or E2BSA. BlockingS‐palmitoylation of ERα with C451A mutation and/or pre‐treatment with 2‐Bromopalmitate did not prevent but instead enhanced E2or E2BSA‐elicited induction of TH promoter activity. These findings reveal, for the first time, that estradiol induction of TH gene transcription with ERα in PC12 cells involves membrane‐initiated estradiol signaling, rapid activation of dual PKA/MEK signaling pathways, leading to CREB phosphorylation, acting at CRE/CaRE. The data demonstrate possible mechanism whereby estradiol affects catecholaminergic systemsin vivo.