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Regulation of mRNA Partitioning to the Endoplasmic Reticulum

mRNA 内质网分配的调节

基本信息

批准号：
7841846
负责人：
Christopher V. Nicchitta
金额：
$ 29.34万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-05-01 至 2012-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7841846
关键词：
5&apos Untranslated Regions Address Binding Biochemical Biological Cell Fractionation Cell Line Cell Survival Cell model Cells Cellular Stress Complex Coupled Cultured Cells Cytosol Data Deletion Mutagenesis Diabetes Mellitus Disease Elements Endoplasmic Reticulum Eukaryotic Cell Family Fractionation Health Histones Homeostasis In Situ In Situ Hybridization In Vitro Initiator Codon Integral Membrane Protein Life Mammalian Cell Mediating Membrane Proteins Messenger RNA Modeling Molecular Mutagenesis Obesity Open Reading Frames Pathway interactions Pattern Peptide Signal Sequences Physiological Plants Polyribosomes Process Protein Biosynthesis Protein translocation Proteins Regulation Regulatory Element Reporter Reporting Research Research Personnel Reticulum Ribosomes Role Sequence Deletion Signal Recognition Particle Signal Transduction Site Stress Stroke Structure Techniques Testing Therapeutic Intervention Translations Uncertainty Untranslated Regions Yeasts abstracting biological adaptation to stress cDNA Arrays defined contribution fly human disease in vivo insight knowledge base member mutant polypeptide programs secretory protein signal recognition particle 72 small hairpin RNA stem tissue culture trafficking

项目摘要

DESCRIPTION (provided by applicant): In contemporary models, eukaryotic cells segregate the synthesis of different classes of proteins; secretory/integral membrane proteins are made on the endoplasmic reticulum (ER) and soluble proteins in the cytosol. Recent studies of mRNA partitioning between the cytosol and ER compartments have, however, identified a prominent role for the ER in the synthesis of both soluble and secretory/membane proteins alike. In addition to describing new functions for the ER in global protein synthesis, these findings identify a significant gap in our understanding of how eukaryotic cells partition mRNAs between the two compartments and in turn regulate the synthesis of these two broad classes of proteins. To address this gap, we are focusing our research efforts to 1) understand how eukaryotic cells partition mRNAs between the cytosol and the endoplasmic reticulum (ER) and 2) determine how eukaryotic cells regulate the protein synthesis activity of the cytosol and ER compartments during homeostasis and cell stress. This research is expected to provide significant insights into the regulatory mechanisms governing protein synthesis in health and disease. In addition, this research will serve as a significant contribution to our understanding of the mRNA localization and protein synthesis pathways that are essential to eukaryotic life. Three specific aims are proposed: i) Define the in vivo role of the SRP pathway in mRNA partitioning to the ER; ii) Identify the mRNA localization signals that confer non-canonical mRNA partitioning to the ER and iii) Define the global role of the ER compartment in cytosolic protein syntehsis. To address these specific aims, we will use mammalian tissue culture cell models and will emphasize established biochemical techniques of cell fractionation, cell biological analysis of reporter mRNA localization in situ and molecular biological analysis of the structure/function elements of mRNAs that confer ER localization. Many prominent human diseases, including obesity, diabetes and stroke activate cell stress responses that profoundly alter the types and amounts of proteins cells synthesize and consequently, cell viability. By understanding how cells decide which and how much of each protein to make, in health and disease, we will understand the basic mechanisms used by cells to respond to pathological stress. By understanding these mechanisms, we hope to identify new targets for therapeutic intervention.

描述（由申请人提供）：在当代模型中，真核细胞分离不同类别蛋白质的合成；分泌/整合膜蛋白在内质网 (ER) 上产生，可溶性蛋白在细胞质中产生。然而，最近对细胞质和内质网区室之间 mRNA 分配的研究发现，内质网在可溶性和分泌/膜蛋白等合成中发挥着重要作用。除了描述内质网在整体蛋白质合成中的新功能之外，这些发现还表明我们对真核细胞如何在两个区室之间分配 mRNA 并进而调节这两大类蛋白质的合成的理解存在重大差距。为了解决这一差距，我们的研究重点是 1) 了解真核细胞如何在细胞质和内质网 (ER) 之间分配 mRNA，2) 确定真核细胞在稳态和细胞应激期间如何调节细胞质和 ER 区室的蛋白质合成活性。这项研究有望为健康和疾病中蛋白质合成的调控机制提供重要见解。此外，这项研究将为我们理解真核生命所必需的 mRNA 定位和蛋白质合成途径做出重大贡献。提出了三个具体目标： i) 定义 SRP 通路在 mRNA 分配到 ER 中的体内作用； ii) 识别赋予 ER 非典型 mRNA 分配的 mRNA 定位信号，以及 iii) 定义 ER 区室在胞质蛋白合成中的整体作用。为了实现这些具体目标，我们将使用哺乳动物组织培养细胞模型，并强调已建立的细胞分级生化技术、报告基因 mRNA 原位定位的细胞生物学分析以及赋予 ER 定位的 mRNA 结构/功能元件的分子生物学分析。许多重要的人类疾病，包括肥胖、糖尿病和中风，都会激活细胞应激反应，从而深刻改变细胞合成蛋白质的类型和数量，从而改变细胞活力。通过了解细胞如何决定在健康和疾病中产生每种蛋白质的种类和数量，我们将了解细胞响应病理应激的基本机制。通过了解这些机制，我们希望找到治疗干预的新靶点。