Mechanism of Actin mRNA Localization and Localized Translation in Neurons

神经元中肌动蛋白 mRNA 定位和定位翻译的机制

基本信息

批准号：
9147647
负责人：
Robert H Singer
金额：
$ 55.96万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE, INC
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-01 至 2017-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9147647
关键词：
3&apos Untranslated Regions Actins Address Adhesions Affect Affinity Behavior Binding Binding Proteins Biological Assay Birth Brain Brain-Derived Neurotrophic Factor Breast cancer metastasis Cell Line Cell physiology Cells Cellular Structures Characteristics Chimeric Proteins Cytoplasmic Granules Defect Dendrites Destinations Detection Development Diabetes Mellitus Disease Distant Drug Addiction Drug resistance Embryo Event Fibroblasts Fluorescence Resonance Energy Transfer Frequencies Funding Generations Genetic Translation Goals Grant Growth Growth Cones Heart Hour Image Immunofluorescence Immunologic Individual Kinetics Knockout Mice Knowledge Label Lead Learning Life Location Maintenance Mammary gland Maps Measures Memory Messenger RNA Metabolism Methodology Methods Microscopy Movement Mus Neurites Neurons Neurotransmitter Receptor Newborn Infant Open Reading Frames Phosphotransferases Photons Potassium Chloride Protein Biosynthesis Protein Family Proteins RNA Reading Regulation Reporter Resistance Ribosomes Role Shapes Signal Transduction Site Stimulus Structure Subfamily lentivirinae Synapses Talin Techniques Technology Time Tissues Trans-Activators Transgenic Animals Transgenic Mice Translating Translational Regulation Translations Travel Vertebral column aptamer base cell growth cell motility design disorder prevention innovation knock-down malignant neurologic neoplasms meter method development mouse model nervous system disorder neurodevelopment new technology novel polypeptide postsynaptic prevent public health relevance research study single molecule small hairpin RNA tool translation assay

项目摘要

DESCRIPTION (provided by applicant): ABSTRACT: The regulation of mRNA is critical to cellular function. Over the years of this grant we discovered that an important mode of regulation is the ability of messenger RNA to become localized at specific regions in cells where synthesis of specific proteins can be spatially compartmentalized. This has profound implications for cell structure and function since all cells have characteristic shapes and structure, which are essential for them to perform their function. The best example of this is the neuron, where the synapses are very far away from the cell body, sometimes meters away. Messenger RNA may have to travel these far distances and then be activated to make proteins upon stimulation of specific synapses at a precise moment. This is the basis of learning and memory. The mechanism by which the mRNA can remain quiescent for long periods of time and then become active upon a specifically localized stimulus is unknown. The Specific Aims of this proposal are directed toward developing new technology to address this question. We have made significant experimental, technical and conceptual advances that have allowed us to observe single molecules of mRNA. For instance we have made a mouse where every molecule of the ß-actin mRNA, which makes an essential protein is labeled. This will allow us to observe these molecules in living cells and tissues. We found that the mRNA travels to distant regions of the cell because of a sequence known as the zipcode. We discovered that this sequence binds a protein, the zipcode binding protein (ZBP1) and this binding silences the mRNA until it reaches its final destination. To activate the mRNA to translate, the protein must be modified at its destination by a kinase that phosphorylates it at a specific site. We believe ZBP1 is the key to understanding the regulatory events that occur at specific locations in the cell, for instance at the synapse, where ß-actin is necessary for stabilizing spines important for their presentation to incoming signals. In support of the importance of this protein, if we delete it in mice, the result is lethal, newborn mice do not survive and their brains show defects in organization of the neuronal layers. We have shown that the protein is essential for proper migration of cells, such as fibroblasts and neurons, and this we believe is due to the ability of the cell to direct the synthesis of actin in a polarized location, where it can polymerize and drive the extension of cell structures involved in movement. ZBP1 is also implicated in disease prevention. After birth, the expression of ZBP1 is repressed, but we have been able to make a transgenic mouse that expresses ZBP1 exogenously in the brain and show that these mice have profoundly altered behavior: they become resistant to drug addiction. Furthermore, expression of ZBP1 in the mammary gland makes the mice resistant to breast cancer metastasis. We have also recently discovered that the ZBP1 family of proteins (there are three) is also implicated in preventing neurological diseases and diabetes. Hence not only will study of this protein reveal how mRNA is regulated, but also how disruption of this regulation can lead to a broad variety of diseases.

描述（由申请人提供）：摘要：mRNA的调节对细胞功能至关重要。在这笔赠款的几年中，我们发现一种重要的调节方式是使者RNA在特定蛋白质合成的细胞中定位在特定区域的能力。这对细胞结构和功能具有深远的影响，因为所有细胞都有特征形状和结构，这对于它们执行其功能至关重要。最好的例子是神经元，那里的突触离细胞体很远，有时会远。 Messenger RNA可能必须在精确时刻刺激特定突触时激活这些遥远的距离，然后被激活以制成蛋白质。这是学习和记忆的基础。 mRNA可以长时间保持静止，然后在特定局部刺激上保持活性的机制是未知的。该提案的具体目的是针对开发新技术来解决这个问题的。我们已经做出了重大的实验，技术和概念进步，使我们能够观察到mRNA的单分子。例如，我们制作了一只小鼠，其中每个分子的ß-肌动蛋白mRNA的每个分子都标记为必不可少的蛋白质。这将使我们能够在活细胞和组织中观察这些分子。我们发现，由于称为Zipcode的序列，mRNA传播以区分细胞区域。我们发现该序列结合了蛋白质，Zipcode结合蛋白（ZBP1），并且该结合使mRNA保持沉默，直到达到最终目的地为止。为了激活mRNA以翻译，必须通过在特定位点磷酸化的激酶在其目的地修饰蛋白质。我们认为，ZBP1是了解细胞中特定位置的调节事件的关键，例如在突触中，ß-肌动蛋白对于稳定刺是对它们向传入信号的呈现很重要的必要条件。为了支持该蛋白质的重要性，如果我们将其删除在小鼠中，则结果是致命的，新生小鼠无法生存，并且它们的大脑在神经元层的组织中显示出缺陷。我们已经表明，该蛋白质对于细胞（例如成纤维细胞和神经元）的适当迁移至关重要，我们认为这是由于细胞将肌动蛋白在极化位置引导合成的能力所致，在极化位置可以聚合并驱动涉及运动中涉及的细胞结构的扩展。 ZBP1也隐含在疾病预防中。出生后，ZBP1的表达得到反映，但是我们能够制作出一种在大脑中外源表达ZBP1的转基因小鼠，并表明这些小鼠的行为有很大改变：它们对吸毒成瘾具有抗性。此外，ZBP1在乳腺中的表达，我们最近还发现，在预防神经系统疾病和糖尿病方面，ZBP1蛋白质（有三种）也暗示。因此，研究这种蛋白质不仅揭示了mRNA的调节方式，而且还会揭示该调节的破坏如何导致多种疾病。