Mechanisms and Functions of Shoc2-Transduced Cellular Signals

Shoc2 转导的细胞信号的机制和功能

• 批准号: 10387097
• 负责人: Emilia Galperin
• 金额: $ 2.51万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10387097
• 关键词: Address; Biological Markers; Cell Adhesion; Cell physiology; Cells; Clinical; Complex; Congenital Abnormality; Development; Disease; Enzymes; Funding; Future; Genes; Genetic; Intervention; Lead; MAPK3 gene; Malignant Neoplasms; Mediating; Microscopy; Modeling; Molecular; Mutation; National Institute of General Medical Sciences; Outcome; Pathway interactions; Patients; Proteins; Research; Scaffolding Protein; Signal Pathway; Signal Transduction; Sorting - Cell Movement; Spatial Distribution; Specificity; Syndrome; Ubiquitin; Vision; Work; Zebrafish; base; biophysical techniques; cell motility; developmental disease; equipment acquisition; in vivo; innovation; new therapeutic target; novel diagnostics; novel therapeutic intervention; programs; public health relevance; scaffold; targeted treatment; therapeutic development; transmission process; tumor; virtual

The overall vision of our NIGMS-supported program is to address the molecular mechanisms of the extracellular signal-regulated kinases 1 and 2 (ERK1/2) signal transmission as guided by scaffold proteins. Essential for the normal functioning of the cell, the ERK1/2 ability to “rewire” signaling pathways is a major problem for clinical intervention. The rationale for the proposed studies is that a thorough understanding of the mechanism by which key regulators, such as scaffold protein Shoc2, modify ERK1/2 signaling is essential for understanding dysregulated mechanisms in ERK1/2-related diseases and for identifying new drug targets. Our studies have raised many exciting questions, and the most transformative research will be pursued in the next five-year period. Our efforts during the first cycle of NIGMS funding led to the discovery that the ubiquitin machinery regulates the signal transmission of ERK1/2 signals through the Shoc2 scaffolding module and controls assembly of the proteins in the complex. We identified key enzymes modulating the ability of Shoc2 to transmit ERK1/2 signals. We then discovered that organized spatial distribution of the Shoc2 complexes is critical for the proper transmission of ERK1/2 signals. Our work further revealed a distinct function of the signals transduced by the Shoc2 scaffold in ERK1/2 in mediating cell adhesion and motility. We also established that loss of Shoc2 in zebrafish has a systemic effect of on early development recapitulating congenital malformations observed in patients with Shoc2 mutations. For MIRA application, we propose to define the mechanisms by which endosomal sorting regulates the ubiquitin-driven remodeling of the Shoc2 complexes and signaling. The key steps that segregate Shoc2 into the distinct intracellular sorting pathways will be delineated. The molecular details of how Shoc2-activated ERK1/2 signals promote early developmental stages will be determined. We will use an innovative comprehensive approach encompassing genetic, state-of-the-art microscopy, as well as cell-based and biophysical methodologies. Of note, we will use zebrafish as a vertebrate model to investigate the in vivo effect of Shoc2 gene editing on the early stages of development since they offer distinct advantages for studies of development, including their transparency, as well as external and rapid development. The proposed research is expected to produce an ambitious and comprehensive mechanistic understanding of how Shoc2 is involved in determining the specificity of ERK1/2 signaling outcomes, and has the potential for high impact by laying the groundwork for future studies on developmental disorders and contributing to the advancement of novel therapeutic strategies and biomarkers.

我们NIGMS支持的计划的总体愿景是解决 细胞外信号调节激酶1和2(ERK1/2)在支架蛋白的引导下进行信号传递。 ERK1/2对细胞的正常功能是必不可少的，ERK1/2重新连接信号通路的能力是一个主要的 临床干预的问题。建议进行研究的理由是，彻底了解 关键调节因子，如支架蛋白Shoc2，修饰ERK1/2信号的机制是 了解ERK1/2相关疾病的失调机制，并用于确定新药靶点。我们的 研究提出了许多令人兴奋的问题，最具变革性的研究将在下一步继续进行 五年期。 我们在NIGMS第一轮资助中的努力发现，泛素机制 通过Shoc2脚手架模块调节ERK1/2信号的信号传输，并控制 蛋白质在复合体中的组装。我们确定了调节Shoc2传输能力的关键酶 ERK1/2信号。然后我们发现Shoc2复合体的有组织的空间分布对于 ERK1/2信号的正确传输。我们的工作进一步揭示了信号的一种独特功能 由ERK1/2中的Shoc2支架介导的细胞黏附和运动。我们还确定了 斑马鱼Shoc2基因缺失对早期发育的全身性影响 在Shoc2突变患者中观察到畸形。 对于MIRA的应用，我们建议定义内体分选调节 泛素驱动的Shoc2复合体和信号的重塑。将Shoc2分离为 不同的细胞内分选途径将被描绘出来。Shoc2-激活的分子细节 ERK1/2信号促进早期发育阶段将被确定。我们将使用一种创新的 全面的方法包括遗传的、最先进的显微镜以及基于细胞和 生物物理学方法。值得注意的是，我们将使用斑马鱼作为脊椎动物的模型来研究体内的影响 Shoc2基因编辑在发育的早期阶段，因为它们为研究 发展，包括其透明度，以及外部和快速发展。拟议的研究 预计将对Shoc2是如何参与的产生一个雄心勃勃的和全面的机械理解 在确定ERK1/2信号转导结果的特异性方面，并可能通过铺设 为未来关于发育障碍的研究奠定基础，并有助于小说的进步 治疗策略和生物标志物。