Mechanisms and functions of Shoc2-transduced cellular signals

我们的NIGMS支持的计划的总体愿景是解决的分子机制， 细胞外信号调节激酶1和2（ERK 1/2）的信号传导的指导下，支架蛋白。 对于细胞的正常功能至关重要，ERK 1/2“重新连接”信号通路的能力是细胞的主要功能。 临床干预的问题。拟议研究的理由是， 关键调节因子如支架蛋白Shoc 2修饰ERK 1/2信号传导的机制对于 了解ERK 1/2相关疾病的失调机制，并确定新的药物靶点。我们 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个世纪继续进行。 五年期。 我们在NIGMS第一轮资助期间的努力导致了泛素机制的发现， 调节ERK 1/2信号通过Shoc 2支架模块的信号传递，并控制 在复合物中组装蛋白质。我们确定了调节Shoc 2传递能力的关键酶， ERK 1/2信号。然后，我们发现Shoc 2复合物的有组织的空间分布对于 ERK 1/2信号的正常传递。我们的工作进一步揭示了信号的独特功能 在介导细胞粘附和运动中，由ERK 1/2中的Shoc 2支架转导。我们还确定， 斑马鱼中Shoc 2的缺失对早期发育具有系统性影响， 在具有Shoc 2突变的患者中观察到畸形。 对于MIRA的应用，我们建议定义内体分选调节细胞增殖的机制。 泛素驱动的Shoc 2复合物和信号转导的重塑。将Shoc 2分离为 将描绘不同的细胞内分选途径。Shoc 2如何激活 ERK 1/2信号促进早期发育阶段将被确定。我们将采用创新的 全面的方法，包括遗传，最先进的显微镜，以及基于细胞的， 生物物理方法学。值得注意的是，我们将使用斑马鱼作为脊椎动物模型来研究体内效应 Shoc 2基因编辑在发育早期阶段的作用，因为它们为研究 发展，包括其透明度，以及外部和快速发展。拟议研究 预计将产生一个雄心勃勃的和全面的机械理解如何Shoc 2参与 在确定ERK 1/2信号转导结果的特异性方面，并通过奠定 为未来发展障碍的研究奠定基础，并为促进新的 治疗策略和生物标志物。

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.