Request to Transfer R01GM105707-03 Novel regulatory mechanisms of Drosophila Pumilio and Nanos

请求转让 R01GM105707-03 果蝇 Pumilio 和 Nanos 的新颖调控机制

• 批准号: 9225476
• 负责人: Aaron Charles Goldstrohm
• 金额: $ 25.86万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-20 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9225476
• 关键词: Request; Transfer; R01GM105707; 03; Novel

DESCRIPTION (provided by applicant): The long term goal of this research is to reveal how gene expression is regulated at the post- transcriptional level. Control of messenger RNA (mRNA) translation and degradation underlies important biological processes including development, fertility and neurological functions. The proposed work focuses on the archetypal mRNA regulator, Pumilio (Pum). Pum is a member of the PUF family of eukaryotic RNA binding proteins. Pum binds an extensive group of messenger RNAs including those that encode key developmental morphogens. Upon binding to an mRNA, Pum represses expression of the encoded protein. The proposed research seeks to discover the mechanism of Pum repression and determine how Pum activity is modulated. We developed novel assays to measure Pum activity in Drosophila cells and discovered multiple new domains that potently repress protein expression. The repressive activity of the Pum repression domains do not rely on previously identified cofactors, indicating novel repression mechanism(s). In the first aim, we measure the impact of the repression domains on translation and mRNA degradation. We seek to discover cofactors necessary for their activity. We then investigate the role of each repression domain in the control of embryonic development. We discovered an autoregulatory switch that controls Pum activity and may be modulated by phosphorylation. In the second aim, we dissect the mechanism of this autoregulatory switch using functional and protein interaction assays. The role of Pum autoregulation during embryogenesis will be explored. This research will determine novel mechanisms of Pum repression and illuminate how that activity is regulated in response to protein modifications. The resulting discoveries are expected to broadly enhance our understanding of post-transcriptional control and specifically improve knowledge of gene regulation in development, fertility, stem cell proliferation, and the nervous system.

描述（由申请人提供）：本研究的长期目标是揭示基因表达是如何在转录后水平调节的。信使RNA（mRNA）翻译和降解的控制是发育、生育和神经功能等重要生物学过程的基础。拟议的工作集中在原型mRNA调节子Pumilio（Pum）上。Pum是真核生物RNA结合蛋白PUF家族的成员。Pum结合了大量的信使RNA，包括那些编码关键发育形态因子的信使RNA。在与mRNA结合后，Pum抑制编码蛋白的表达。这项研究旨在发现Pum抑制的机制，并确定Pum活性是如何调节的。我们开发了新的检测方法来测量果蝇细胞中的Pum活性，并发现了多个有效抑制蛋白质表达的新结构域。Pum阻遏结构域的阻遏活性不依赖于先前鉴定的辅因子，表明新的阻遏机制。在第一个目标中，我们测量阻遏结构域对翻译和mRNA降解的影响。我们试图发现其活动所必需的辅助因子。然后，我们调查的作用，每个抑制域在胚胎发育的控制。我们发现了一个自动调节开关，控制Pum的活动，并可能通过磷酸化调节。在第二个目标中，我们使用功能和蛋白质相互作用测定来剖析这种自动调节开关的机制。Pum的自动调节在胚胎发生过程中的作用将被探讨。这项研究将确定Pum抑制的新机制，并阐明该活性是如何响应蛋白质修饰而调节的。由此产生的发现有望广泛增强我们对转录后控制的理解，并特别提高发育，生育，干细胞增殖和神经系统中基因调控的知识。