Administrative Supplement for Control and Function of Ndr/LATS Signaling Systems (GM084223)

Ndr/LATS 信号系统控制和功能的行政补充（GM084223）

• 批准号: 9137401
• 负责人: ERIC Lyle WEISS
• 金额: $ 8.01万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9137401
• 关键词: Administrative Supplement; Architecture; Behavior; Binding; Binding Proteins; Binding Sites; Biochemical; Biological; Biological Assay; C-terminal; Catalytic Domain; Cell Polarity; Cell Proliferation; Cell Proliferation Regulation; Cell Separation; Cell division; Cell physiology; Cells; Collaborations; Complex; Congenital Abnormality; Consensus; Cytokinesis; Data; Docking; Drosophila genus; Ensure; Eukaryotic Cell; Event; Gene Expression; Genes; Genetic Transcription; Genetic Translation; Goals; Growth; Health; Human; Individual; Life; Link; Maintenance; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Mediating; Messenger RNA; Mitotic; Mitotic spindle; Molecular Biology; Morphogenesis; Neurons; Normal tissue morphology; Orthologous Gene; Pathway Analysis; Pathway interactions; Peptides; Phage Display; Phosphorylation; Phosphotransferases; Principal Investigator; Process; Proteins; Recruitment Activity; Regulation; Saccharomycetales; Shapes; Signal Pathway; Signal Transduction; Site; Specificity; Speed; Structure; Substrate Specificity; System; Testing; Time; Translations; Universities; Work; Yeasts; base; cell behavior; cell cortex; cell growth; crosslink; in vivo; interest; novel; polarized cell; programs; rapid growth; transcription factor

DESCRIPTION (provided by applicant): Signaling pathways in which GCK group "Mst/hippo" kinases regulate AGC group "Ndr/LATS" kinases are ancient controllers of growth, proliferation, and architecture of eukaryotic cells. Our broad goal is to define the diverse intracellular processes these pathways regulate and determine the mechanisms underlying this control. Forms of Mst/hippo signaling ("hippo-warts" pathways) that suppress metazoan cell proliferation in metazoans by inhibiting YAP/yorkie-related transcriptional co-activators have been the subject of intensive recent interest. However, the distinct and highly conserved "hippo-trc" form of this pathway in which large "furry" related proteins mediate Mst/hippo kinase activation of Ndr/tricornered kinases have dramatically different functions; comparatively little is known about them, and they are the focus of this project. The hippo-trc pathways are important for polarized growth and organization of cellular extensions, neuron morphogenesis, mitotic spindle organization, and positive regulation of cell proliferation. We have successfully studied the system in budding yeast, which use a conserved hippo-trc pathway known as the "RAM network" to control cell division and polarized growth. Under close regulation by mitotic exit machinery, this pathway directly drives asymmetric localization and activity of a transcription factor that turns on expression of genes involved in the final step of cytokinesis. In addition to this primordial cell fate decision, budding yeast hippo-trc signaling promotes maintenance of cell polarity and regulates translation of proteins required for physical expansion of the cell during rapid growth. This project aims to define the regulatory mechanisms and downstream targets of Cbk1. Through combined computational and experimental work we have discovered that a novel "docking motif" peptide recruits this Ndr/LATS kinase to in vivo substrates through interaction with the kinase catalytic domain. We have crystallized the Mob2-Cbk1 complex and solved its structure, the first for any Ndr/LATS kinase, and will use this information to guide analysis of the kinase's activation mechanisms. We will define how the docking motif binds to Cbk1's kinase domain, analyze effects caused by disruption of this interaction in vivo, and determine if the novel substrate docking behavior we have discovered in budding yeast also occurs with metazoan orthologs. When combined with existing interaction and phosphoproteomic data, our analysis of substrate docking and consensus motif conservation at least triples the number of high confidence Cbk1 targets. In addition to extending our analysis of the pathway's regulation of mRNA translation, we will explore this expanded regulatory network to gain a more comprehensive mechanistic understanding of this hippo-trc pathway's control of cell division and morphogenesis.

描述（由申请人提供）：GCK组“Mst/hippo”激酶调节AGC组“Ndr/LATS”激酶的信号传导途径是真核细胞生长、增殖和结构的古老控制器。我们的总体目标是定义这些途径调节的不同细胞内过程并确定这种控制的机制。通过抑制 YAP/yorkie 相关转录共激活因子来抑制后生动物中后生动物细胞增殖的 Mst/hippo 信号传导（“hippo-warts”途径）的形式已成为最近人们强烈关注的主题。然而，该途径独特且高度保守的“hippo-trc”形式（其中大的“毛茸茸”相关蛋白介导 Ndr/三角激酶的 Mst/hippo 激酶激活）具有显着不同的功能；人们对它们知之甚少，但它们是该项目的重点。 hippo-trc 通路对于细胞延伸的极化生长和组织、神经元形态发生、有丝分裂纺锤体组织和细胞增殖的正向调节非常重要。我们已经成功地研究了芽殖酵母中的系统，该系统使用被称为“RAM网络”的保守hippo-trc途径来控制细胞分裂和极化生长。在有丝分裂退出机制的密切调控下，该途径直接驱动转录因子的不对称定位和活性，从而开启参与胞质分裂最后步骤的基因的表达。除了这种原始细胞命运决定之外，出芽酵母 hippo-trc 信号传导还能促进细胞极性的维持，并调节细胞在快速生长过程中物理扩张所需的蛋白质翻译。该项目旨在明确Cbk1的监管机制和下游目标。通过计算和实验相结合的工作，我们发现一种新的“对接基序”肽通过与激酶催化结构域的相互作用将这种 Ndr/LATS 激酶募集到体内底物上。我们已经结晶了 Mob2-Cbk1 复合物并解析了其结构，这对于任何 Ndr/LATS 激酶来说都是首次，并将利用这些信息来指导激酶激活机制的分析。我们将定义对接基序如何与 Cbk1 的激酶结构域结合，分析体内这种相互作用破坏所引起的影响，并确定我们在芽殖酵母中发现的新底物对接行为是否也发生在后生动物直向同源物中。当与现有的相互作用和磷酸化蛋白质组数据相结合时，我们对底物对接和共有基序保守性的分析至少使高置信度 Cbk1 靶点的数量增加了三倍。除了扩展我们对该通路对 mRNA 翻译的调节的分析之外，我们还将探索这个扩展的调节网络，以获得对 hippo-trc 通路对细胞分裂和形态发生的控制的更全面的机制理解。