ROLE OF SHOC2 PROTEIN IN SPATIO-TEMPORAL REGULATION OF ERK KINASE CASCADE

SHOC2 蛋白在 ERK 激酶级联时空调控中的作用

• 批准号: 8360579
• 负责人: Emilia Galperin
• 金额: $ 25.82万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8360579
• 关键词: Biochemical; Biological Assay; Centers of Research Excellence; Complex; Endocytosis; Endosomes; Epidermal Growth Factor Receptor; Event; Fluorescence Microscopy; Funding; Goals; Grant; MAP Kinase Gene; MAPK Signaling Pathway Pathway; MAPK3 gene; MEKs; Methodology; Methods; Molecular; National Center for Research Resources; Phosphotransferases; Principal Investigator; Proteins; Reagent; Regulation; Reporting; Research; Research Infrastructure; Resources; Role; Scaffolding Protein; Signal Transduction; Source; Specificity; United States National Institutes of Health; base; cancer cell; cost; human disease; member; neoplastic cell; reconstitution; trafficking

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Activation of Epidermal Growth Factor (EGF) Receptor triggers a network of signal transduction events including activating the MAPK cascade along with accelerating endocytosis of EGF-receptor complexes. MEK and ERK proteins are localized on endosomes, however the precise mechanism and the role of endosomal targeting are yet to be understood. In addition, members of the Ras-RAF-MEK-ERK cascade are reported to be organized into signal-transducing modules regulated by scaffold proteins. The suggested role of scaffold proteins is to enhance the specificity and the selectivity of interactions by bringing kinase-substrate partners into these modules. We seek to elucidate endocytosis involvement in regulating of the MAPK signaling pathway by the scaffold protein Shoc2. Shoc2 is a positive regulator of ERK1/2 activation and signaling. We demonstrated that Shoc2 relocates to a subset of endosomes upon EGFR activation. This protein is unique in its ability to control MAPK activation that is also regulated by endocytic trafficking. However, the mechanism is poorly understood. Essentially, our goal will be to define the functional importance of endosomal localization of Shoc2 for MAPK activity by utilizing various biochemical assays and advanced methods of fluorescence microscopy. We will analyze the mechanism of the spatial regulation of Shoc2 and reconstitute Shoc2 in cancer cells to study the contribution of Shoc2 to MAPK activation in tumor cells. A number of reagents and newly developed methodologies will allow us to pursue this goal.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 表皮生长因子 (EGF) 受体的激活会触发信号转导事件网络，包括激活 MAPK 级联以及加速 EGF-受体复合物的内吞作用。 MEK 和 ERK 蛋白定位在内体上，但内体靶向的精确机制和作用尚不清楚。此外，据报道 Ras-RAF-MEK-ERK 级联的成员被组织成由支架蛋白调节的信号转导模块。支架蛋白的作用是通过将激酶-底物伙伴带入这些模块来增强相互作用的特异性和选择性。我们试图阐明内吞作用参与支架蛋白 Shoc2 对 MAPK 信号通路的调节。 Shoc2 是 ERK1/2 激活和信号转导的正调节因子。我们证明 Shoc2 在 EGFR 激活后重新定位到内体的一个子集。该蛋白的独特之处在于其控制 MAPK 激活的能力，而 MAPK 激活也受到内吞运输的调节。然而，人们对该机制知之甚少。本质上，我们的目标是通过利用各种生化测定和先进的荧光显微镜方法来确定 Shoc2 内体定位对于 MAPK 活性的功能重要性。我们将分析Shoc2的空间调控机制，并在癌细胞中重建Shoc2，以研究Shoc2对肿瘤细胞中MAPK激活的贡献。许多试剂和新开发的方法将使我们能够实现这一目标。