Control and Function of Ndr/LATS Signaling Systems

Ndr/LATS 信号系统的控制和功能

• 批准号: 9020232
• 负责人: ERIC Lyle WEISS
• 金额: $ 28.58万
• 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/9020232
• 关键词: 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; Transcription Coactivator; Translations; Universities; Work; Yeasts; base; cell behavior; cell cortex; cell growth; crosslink; in vivo; interest; novel; phosphoproteomics; 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的生长，增殖和结构的古代控制器。我们的广泛目标是定义这些途径调节的各种细胞内过程，并确定该控制的机制。 MST/HIPPO信号传导的形式（“河马”途径）通过抑制与YAP/Yorkie相关的转录共激活因子来抑制后生动物中的后生细胞增殖。然而，这种途径的独特且高度保守的“河马trc”形式在这种途径中，大型“毛茸茸”相关蛋白介导了NDR/Tricornered激酶的MST/HIPPO激酶激活具有巨大的功能。对它们的了解很少，它们是该项目的重点。河马-TRC途径对于细胞延伸，神经元形态发生，有丝分裂的纺锤体组织和细胞增殖的阳性调节很重要。我们已经成功地研究了萌芽酵母的系统，该系统使用保守的河马-TRC途径称为“ RAM网络”来控制细胞分裂和偏振生长。在有丝分裂出口机械的密切调控下，该途径直接驱动转录因子的不对称定位和活性，该途径打开了涉及细胞球运动的基因的表达。除了这种原始细胞命运的决策外，萌芽的酵母hippo-trc信号促进了细胞极性的维持，并调节快速生长过程中细胞物理膨胀所需的蛋白质的翻译。该项目旨在定义CBK1的调节机制和下游目标。通过合并的计算和实验工作，我们发现一种新颖的“对接基序”肽通过与激酶催化域的相互作用来募集这种NDR/LATS激酶以体内底物。我们已经结晶了MOB2-CBK1复合物，并解决了其结构，这是任何NDR/LATS激酶的第一个结构，并将使用此信息来指导激酶的激活机制分析。我们将定义对接基序如何与CBK1的激酶结构域结合，分析由于体内这种相互作用的破坏而引起的效果，并确定我们在萌芽酵母中发现的新型底物对接行为是否也发生在萌芽的酵母中。当与现有的相互作用和磷蛋白质组学数据结合使用时，我们对底物对接和共识基序保护的分析至少三倍，高置信度CBK1靶标的数量。除了扩展我们对途径对mRNA翻译的调节的分析外，我们还将探索这个扩展的调节网络，以获得对这一HIPPO-TRC途径对细胞分裂和形态发生的控制的更全面的机械理解。