Characterization of MWS/DEN, a novel regulator of amino acid signaling to mTORC1

MWS/DEN 的表征，一种新型 mTORC1 氨基酸信号传导调节剂

基本信息

批准号：
8722844
负责人：
Lynne Chantranupong
金额：
$ 4.27万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-01 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8722844
关键词：
Adverse effects Affect Amino Acids Autophagocytosis Biogenesis Biological Assay Biological Markers Breathing Catabolic Process Cell Culture Techniques Cells Clinical Clinical Trials Complex Cues Databases Development Diabetes Mellitus Energy Supply Exhibits FDA approved Frequencies GTPase-Activating Proteins Gene Silencing Genes Genetic Translation Genomics Goals Growth Growth Factor Guanosine Triphosphate Guanosine Triphosphate Phosphohydrolases Hydrolysis Immunofluorescence Immunologic Immunoprecipitation Institutes Insulin Knowledge Lead Malignant Neoplasms Manuscripts Mass Spectrum Analysis Mediating Molecular Mutation Nerve Degeneration Normal Cell Nutrient Pathway interactions Phosphotransferases Process Protein Kinase Protein Subunits Proteins Proteomics Regulation Renal Cell Carcinoma Reporting Ribosomes Role Signal Transduction Sirolimus Stimulus Stress Surface Techniques Therapeutic Translations Transplantation Tumor Suppressor Proteins Ubiquitination Work cancer cell carcinogenesis genetic analysis human disease in vivo inhibitor/antagonist insight loss of function novel novel therapeutics protein complex protein purification public health relevance response sarcoma scaffold stem tumor ubiquitin ligase

项目摘要

DESCRIPTION (provided by applicant): The mechanistic target of rapamycin complex 1 (mTORC1) protein kinase is a master growth regulator. It integrates inputs from growth factors as well as nutrient and energy supplies to control key biosynthetic and catabolic processes, including protein translation, ribosome biogenesis and autophagy. As might be expected, this pathway is deregulated in a number of human diseases, ranging from neurodegeneration to cancer. Amongst the diverse inputs that impinge on mTORC1, amino acids are unique in that they are absolutely required for kinase activity, despite the presence of multiple activating stimuli. Amino acids utilize an independent mechanism from other cues to regulate mTORC1; however, this mechanism remains poorly characterized despite recent progress. By gaining a more complete understanding of how amino acids signal to mTORC1, we may also gain insight into more effective targets for pharmacological inhibition of mTORC1 with minimized side effects. We have used an unbiased proteomic approach to identify a novel octameric complex consisting of two interacting subcomplexes. The first subcomplex, MWS, is a positive regulator of the amino acid branch and contains Mios, WDR24, WDR59, Sec13, and Seh1L. The second subcomplex, DEN, contains DEPDC5, Nprl2, and Nprl3 and is the first negative regulator of the amino acid branch of mTORC1 to be identified. DEN's inhibitory role stems from its function as a GTPase activating protein (GAP) for the RagA/B GTPases, a key activator of the mTORC1 pathway. Importantly, as a negative regulator, DEN has the potential to be deregulated in cancer as a tumor suppressor. The goal of this project is to define and characterize the mechanisms of DEN and MWS in amino acid signaling to mTORC1 and to interrogate the possible role of DEN in carcinogenesis. To achieve this goal, we propose the following aims: 1. Further characterize the GAP activity of DEN towards RagA/B. 2. Determine the mechanism by which MWS regulates DEN in response to amino acids by exploring localization and a potential ubiquitin ligase function for MWS. 3. Interrogate the role for DEN as a tumor suppressor complex in cancer. Through a combination of hypothesis driven and unbiased approaches, our work will clarify how the amino acid pathway of mTORC1 functions in normal cells and how it can be deregulated in carcinogenesis. These insights may lead to the development of novel biomarkers and therapeutic strategies for cancer.

描述（由申请人提供）：雷帕霉素复合物1（MTORC1）蛋白激酶的机械靶标是主要生长调节剂。它整合了来自生长因子以及养分和能源供应的投入，以控制关键的生物合成和分解代谢过程，包括蛋白质翻译，核糖体生物发生和自噬。可以预料的是，该途径在许多人类疾病中被放松，从神经变性到癌症。在撞击MTORC1上的多种输入中，氨基酸是独一无二的，因为尽管存在多种激活刺激，但它们绝对需要激酶活性。氨基酸利用来自其他提示的独立机制来调节MTORC1；但是，尽管进展最近，这种机制的特征仍然很差。通过对氨基酸如何向MTORC1发出信号的完整了解，我们还可以深入了解更有效的MTORC1药理抑制靶标具有最小的副作用。我们已经使用了一种无偏的蛋白质组学方法来识别由两个相互作用的子复合物组成的新型八聚体络合物。第一个子复合物MWS是氨基酸分支的正调节剂，并包含MIOS，WDR24，WDR59，SEC13和SEH1L。第二个子复合den包含DEPDC5，NPRL2和NPRL3，是MTORC1的氨基酸分支的第一个负调节剂。 DEN的抑制作用源于其作为MTORC1途径的关键激活因子Raga/B GTPases的GTPase激活蛋白（GAP）的功能。重要的是，作为负调节剂，DEN有可能在癌症中以肿瘤抑制剂的形式放松管制。该项目的目的是定义和表征氨基酸信号中DEN和MWS的机制，并询问DEN在致癌作用中的可能作用。为了实现这一目标，我们提出以下目的：1。进一步描述Den对Raga/b的间隙活动。 2。通过探索MWS的定位和潜在的泛素连接酶功能来确定MWS对氨基酸响应DEN的机制。 3。询问DEN作为癌症中肿瘤抑制剂复合物的作用。通过假设驱动和无偏的方法的结合，我们的工作将阐明MTORC1的氨基酸途径如何在正常细胞中的功能以及如何在致癌作用中消失。这些见解可能导致新的生物标志物和癌症治疗策略的发展。