Diversity Supplement - Deciphering the Molecular Mechanisms Underlying Active Scaffolding

 描述(申请人提供)：细胞外信号调节激酶1和2(ERK1/2)信号通路在胚胎发育和肿瘤进展的几个关键步骤中发挥重要作用。它还控制着重要的细胞功能，如细胞凋亡、运动和分化。这一途径的激酶和磷酸酶已被广泛研究，并被靶向治疗。然而，决定ERK1/2信号特异性和协调ERK1/2信号不同生物学结果的机制仍然知之甚少。支架蛋白是ERK1/2信号通路中的关键成员，被认为整合传入信号并传递信号特异性，但它们在信号传播中的作用和作用机制尚不清楚。我们研究的长期目标是揭示支架蛋白如何参与ERK1/2级联调控的生物过程。通过这种方式，可以开发新的治疗策略，以更具体地针对这种级联反应，而不影响其他基本的生物学功能。这项建议的目的是确定关键支架蛋白Shoc2调节ERK1/2介导的细胞运动的潜在机制。Shoc2是胚胎发育所必需的，也是ERK1/2活性的关键调节因子。我们已经证明，斑马鱼中Shoc2的缺失会导致造血功能的缺陷。我们还发现Shoc2整合了内吞和泛素机制来调节ERK1/2信号转导。基于这一初步数据，该提议的中心假设是Shoc2创建了一个信号中枢，以时空方式调节ERK的发育信号。我们的假设将通过追求三个具体目标来检验：目标1将决定Shoc2支架复合体的时空组织和动力学。目的2揭示E3连接酶HUWE1控制Shoc2功能和组装Shoc2支架复合体的分子机制。目的3将确定通过调控胚胎发育的Shoc2支架复合体介导的信号事件。因此，拟议的目标有望提供对Shoc2如何参与决定ERK1/2信号转导结果的特异性的详细理解。这些研究将使用最先进的创新显微镜、遗传学、分子和细胞技术，并将为我们理解支架蛋白对信号的时空控制提供关键的见解。这项研究对于促进发育障碍和癌症进展的新的治疗策略和生物标志物的创新具有重要意义。因此，拟议的研究与NIH的使命相关，即增加对生物过程的了解，并为疾病诊断、治疗和预防方面的进展奠定基础。

 DESCRIPTION (provided by applicant): The extracellular signal-regulated kinases 1 and 2 (ERK1/2) signaling pathway plays an essential role in several critical steps of embryonic development and tumor progression. It also controls critical cellular functions such as apoptosis, motility and differentiation. Kinases and phosphatases of this pathway have been extensively studied and targeted therapeutically. However, the mechanisms that determine the signal specificity and orchestrate the diverse biological outcomes of ERK1/2 signaling are still poorly understood. Scaffold proteins are key players in the ERK1/2 signaling pathway that are thought to integrate incoming signals and deliver signaling specificity, and yet their role in signal propagation and the mechanisms of their action are still unknown. The long-term goal of our research is to reveal how scaffold proteins are involved in the biological processes regulated by the ERK1/2 cascade. In this way, new therapeutic strategies can be developed to more specifically target this cascade without affecting other essential biological functions. The objective of this proposal is to determine the mechanisms underlying the ability of the key scaffold protein, Shoc2, to regulate ERK1/2-mediated cell motility. Shoc2 is essential for embryonic development and a critical regulator of ERK1/2 activity. We have shown that depletion of Shoc2 in zebrafish results in defects in hematopoiesis. We also found that Shoc2 integrates endocytic and ubiquitin machineries to regulate ERK1/2 signaling. Based on this preliminary data, the central hypothesis of this proposal is that Shoc2 creates a signaling hub that regulates ERK's developmental signals in a spatio-temporal manner. Our hypothesis will be tested by pursuing three specific aims: Aim 1 will determine the spatio-temporal organization and dynamics of Shoc2 scaffold complexes. Aim 2 will reveal the molecular mechanism by which the E3 ligase HUWE1 controls Shoc2 function and assembly of the Shoc2 scaffold complexes. Aim 3 will determine the signaling events mediated through the Shoc2 scaffold complexes that control embryonic development. The proposed Aims therefore are expected to provide a detailed understanding of how Shoc2 is involved in determining the specificity of ERK1/2 signaling outcomes. These studies will employ state-of-the-art innovative microscopy, genetic, molecular, and cellular techniques and will provide critical insights into our understanding of the spatial-temporal control of signaling by scaffold proteins. This research has significance for contributing to the advancement of novel therapeutic strategies and biomarker innovations for developmental disorders and cancer progression. Thus, the proposed studies are relevant to the NIH's mission to increase understanding of biological processes and to lay the foundation for advances in disease diagnosis, treatment and prevention.