DEVELOPMENTAL CONTROL OF GNRH EXPRESSION BY POU PROTEINS

POU 蛋白对 GNRH 表达的发育控制

• 批准号: 6381057
• 负责人: Margaret E Wierman
• 金额: $ 16.4万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6381057
• 关键词: gel mobility shift assay; gene expression; genetic mapping; genetic promoter element; genetic regulation; genetically modified animals; gonadotropin releasing factor; laboratory mouse; molecular cloning; neurons; phenotype; protein protein interaction; tissue /cell culture; transcription factor; transfection; western blottings; yeast two hybrid system

DESCRIPTION (Adapted from the Investigator's Abstract): POU homeodomain proteins are transcription factors that play a role in differentiation, and the developmental control of neuroendocrine genes. The GnRH gene is expressed in a subset of hypothalamic neurons and is unique in its plasticity of its phenotype across development. GnRH neuronal phenotype is acquired rapidly as cells complete an early proliferation during embryogenesis, and then undergo loss and reinstatement during the estrous cycle. This led to the question whether POU proteins play a role in the control of neuronal GnRH gene expression. The POU-domain family members, SCIP and Brn-4 were cloned from GT1-7 neuronal cells. Physiological significance was shown by colocalization of SCIP or Brn-4 with GnRH in neurons in vivo. Overproduction of SCIP and Brn-4 was shown to repress or activate GnRH promoter activity, respectively, requiring both direct binding to DNA and protein-protein interactions. This proposal has 4 specific aims: Aim 1-To assess the expression of SCIP and Brn-4 in GnRH neurons in vivo; Aim 2-define the relevance of SCIP modulation of GnRH by targeted overexpression and antagonism of SCIP in transgenic animal; Aim 3-precisely localize the cis-acting elements of the GnRH promoter that mediate repression by SCIP and activation by Brn-4 and their protein partners; 4-identify and clone the protein partners that direct SCIP and Brn-4 action on GnRH. Together these studies will define the physiological role and cellular mechanisms of POU protein control and GnRH gene expression.

描述（改编自研究者的摘要）：POU 同源域 蛋白质是在分化中发挥作用的转录因子，并且 神经内分泌基因的发育控制。 GnRH 基因是 在下丘脑神经元的一个子集中表达，其独特之处在于 其表型在整个发育过程中的可塑性。 GnRH 神经元表型是 当细胞完成早期增殖时迅速获得 胚胎发生，然后在发情期间经历损失和恢复 循环。 这引发了一个问题：POU 蛋白是否在 控制神经元 GnRH 基因表达。 POU域家族成员， SCIP 和 Brn-4 是从 GT1-7 神经元细胞中克隆的。 生理 SCIP 或 Brn-4 与 GnRH 的共定位显示了显着性 体内的神经元。 SCIP 和 Brn-4 的过量生产被证明可以抑制或 分别激活 GnRH 启动子活性，需要两者直接结合 DNA 和蛋白质-蛋白质相互作用。 该提案有 4 个具体目标： 目的1-评估体内GnRH神经元中SCIP和Brn-4的表达； 目标 2-通过靶向定义 SCIP 调节 GnRH 的相关性 SCIP在转基因动物中的过度表达和拮抗作用；瞄准3-精确 定位介导 GnRH 启动子的顺式作用元件 SCIP 的抑制以及 Brn-4 及其蛋白伴侣的激活； 4-识别并克隆指导 SCIP 和 Brn-4 作用的蛋白质伴侣 关于 GnRH。 这些研究将共同​​确定生理作用和 POU 蛋白控制和 GnRH 基因表达的细胞机制。