PROTEIN INTERACTIONS REGULATING PITUITARY SIGNALING

调节垂体信号传导的蛋白质相互作用

• 批准号: 6489618
• 负责人: DAWN L DUVAL
• 金额: $ 9.37万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-02-15 至 2003-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6489618
• 关键词: gene induction /repression; genetic mapping; liquid chromatography; mass spectrometry; prolactin; protein protein interaction; protein structure function; site directed mutagenesis; transcription factor

DESCRIPTION (taken from the application) Transcriptional activity is dependent not only on the binding of transcription factors to cis-acting elements, but also on the ability of those factors to recruit a complex hierarchy of proteins to stabilize the basal transcriptional machinery. Basal lactotroph specific expression of the rat prolactin (rPRL) promoter is dependent on the interaction of the proto-oncoprotein c-Ets-1, a member of the -Ets family of transcription factors, with the pituitary-specific transcription factor, Pit-1, at a composite Ets/Pit-1 DNA binding element in the rPRL promoter. While each of these factors alone is capable of activating rPRL gene transcription, the combination of these two transcription factors results in a marked synergistic response. Although the precise mechanism for this synergy remains unknown, it is clear that physical interaction of these two factors is required. Recent studies suggest that the activity of Pit-1 and Ets-1 may be determined by the formation of co-regulatory complexes. Thus, I hypothesize that the Pit-1/Ets-1 complex mediates the synergistic response of the rPRL promoter by recruiting specific transcription regulatory proteins. The goals of this study are to map the specific amino acids of Pit-1 and Ets-1 responsible for their physical and functional interaction, and to use liquid chromatography/mass spectrometry methods to identify the protein components that are recruited to the Ets-1/Pit-1 complex. Therefore, these studies are likely to provide critical insights into the molecular mechanisms underlying combinatorial factor interactions and how they mediate synergistic responses. Finally, these studies are important to my career development, as they will allow me to gain expertise in the cutting edge field of proteomics and protein structure-function relationships, which I can then utilize to establish my career as an independent investigator.

描述（取自应用程序） 转录活性不仅依赖于转录的结合， 顺式作用元件的因子，而且这些因子的能力， 招募一个复杂的蛋白质层次结构来稳定基础转录 机械.大鼠催乳素（rPRL）的基础催乳素特异性表达 启动子依赖于原癌蛋白c-Ets-1的相互作用， Ets转录因子家族成员，具有垂体特异性 转录因子，Pit-1，在复合Ets/Pit-1 DNA结合元件中， rPRL启动子。虽然这些因素中的每一个都能够单独激活 rPRL基因的转录，这两种转录因子的结合 导致显著的协同反应。虽然精确的机制， 这种协同作用仍然未知，很明显，这些物质的物理相互作用 需要两个因素。最近的研究表明，Pit-1和 Ets-1可以通过共调节复合物的形成来确定。所以我 假设Pit-1/Ets-1复合物介导了 rPRL启动子通过募集特异性转录调节蛋白。的 本研究的目标是绘制Pit-1和Ets-1的特定氨基酸图谱 负责它们的物理和功能相互作用，并使用液体 色谱/质谱法鉴定蛋白质组分 被招募到Ets-1/Pit-1复合体因此，这些研究 可能会提供关键的见解的分子机制， 组合因子相互作用以及它们如何介导协同反应。 最后，这些研究对我的职业发展很重要，因为它们将 让我在蛋白质组学和蛋白质的前沿领域获得专业知识， 结构-功能关系，然后我可以利用它来建立我的 作为一名独立调查员。