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Polarity complexes in cellular differentiation

细胞分化中的极性复合物

基本信息

批准号：
RGPIN-2017-05146
负责人：
Fawcett, James
金额：
$ 2.04万
依托单位：
Dalhousie University
依托单位国家：
加拿大
项目类别：
Discovery Grants Program - Individual
财政年份：
2018
资助国家：
加拿大
起止时间：
2018-01-01 至 2019-12-31
项目状态：
已结题

来源：
https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=650533
关键词：
Polarity complexes cellular differentiation

项目摘要

My NSERC Discovery program has focused on identifying protein-protein complexes that regulate cell polarity. In particular we have been focusing on the role of these complexes play in the development of the synapse, one of the more polarized structures in the CNS. Synapses are sites of cell-cell contact specialized for the transmission of chemical and electrical signals between neurons. The specialized function of the synapse results from the asymmetric distribution of proteins forming the presynaptic and postsynaptic compartments, and involves the coordinated regulation of actin dynamics. Not surprisingly then, signalling pathways that regulate the activity of the small Rho GTPases, Rac, Rho and cdc42, have been shown to play an important role in the development and maintenance of the synapse and synaptic morphology. ******Work from our group has shown that a known polarity protein, Scribble, associates with the adaptor protein nitric oxide synthase adaptor protein (NOS1AP) through a unique protein-protein interaction. This interaction controls synaptic morphology by affecting the activated state of the small RhoGTPase protein, Rac1. To further characterize the role of NOS1AP in the synapse, we conducted a mass spectrometry screen and identified an interaction with N-Cadherin and other proteins implicated in endocytosis. In addition to NOS1AP, we identified that the scaffolding protein, Angiomotin (AMOT), associates with known polarity proteins. We have characterized a role for AMOT and the Lats1 kinase in regulating synaptic morphology. We continue to build on our NSERC program studying the role for known polarity proteins in the synapse. In this granting period we propose to:******i)Characterize a role for NOS1AP in N-cadherin stability in synaptic membranes. In an attempt to further define a role for NOS1AP in the synapse, we conducted a proteomic screen and have now linked NOS1AP with N-cadherin and proteins implicated in the endocytosis of receptor complexes. We will characterize the molecular nature of this NOS1AP complex and determine how this complex regulates the endocytosis of N-Cadherin to affect dendritic spine morphology. ***ii) Characterize a role for Hippo dependent kinase in synaptic morphology. Our previous work has revealed the importance of AMOT in stabilizing actin in dendritic spines, and the importance of the core Hippo kinase Lats1 in this process. We will extend this work to examine the effects of other core Hippo kinases in this process, and whether neuronal activity affects the activity of these core Hippo kinases to then affect dendritic spine morphology.******Together, this program will contribute to our understanding of how known polarity proteins and polarity complexes regulate synaptic morphology and development, and further our understanding of the importance of the evolutionary conserved Hippo pathway in regulating synaptic morphology.**

我的NSERC发现计划专注于识别调节细胞极性的蛋白质-蛋白质复合体。特别是，我们一直在关注这些复合体在突触发育中所起的作用，突触是中枢神经系统中极化程度较高的结构之一。突触是细胞-细胞接触的场所，专门用于神经元之间的化学和电信号传输。突触的特殊功能源于形成突触前和突触后隔区的蛋白质的不对称分布，并涉及到肌动蛋白动力学的协调调节。不足为奇的是，调节小Rho GTP酶Rac、Rho和CDc42活性的信号通路已被证明在突触和突触形态的发育和维持中发挥重要作用。*我们小组的工作表明，一种已知的极性蛋白Scribble通过一种独特的蛋白质-蛋白质相互作用与接头蛋白一氧化氮合酶适配器蛋白(NOS1AP)结合。这种相互作用通过影响小RhoGTP酶蛋白rac1的激活状态来控制突触的形态。为了进一步确定NOS1AP在突触中的作用，我们进行了质谱学筛选，并确定了与N-钙粘附素和其他与内吞作用有关的蛋白质的相互作用。除了NOS1AP，我们还发现支架蛋白Angiomotin(AMOT)与已知的极性蛋白有关。我们已经确定了AMOT和Lats1激酶在调节突触形态中的作用。我们继续在我们的NSERC计划的基础上研究已知的极性蛋白在突触中的作用。在这个授予期间，我们建议：*i)表征NOS1AP在突触膜N-钙粘蛋白稳定性中的作用。为了进一步确定NOS1AP在突触中的作用，我们进行了蛋白质组筛选，现在将NOS1AP与N-钙粘素和参与受体复合体内吞作用的蛋白质联系起来。我们将表征这种NOS1AP复合体的分子性质，并确定该复合体如何调节N-钙粘连蛋白的内吞作用，从而影响树突棘的形态。*ii)表征河马依赖激酶在突触形态中的作用。我们以前的工作揭示了AMOT在稳定树突棘中的肌动蛋白方面的重要性，以及核心河马蛋白激酶Lats1在这一过程中的重要性。我们将进一步研究其他核心河马蛋白在这一过程中的作用，以及神经元活动是否影响这些核心河马蛋白的活性，从而影响树突棘的形态。*这一程序将有助于我们理解已知的极性蛋白质和极性复合体如何调节突触形态和发育，并进一步理解进化保守的河马通路在调节突触形态中的重要性。