Cytokinesis staging mechanisms

细胞分裂分期机制

• 批准号: 10163221
• 负责人: ERIC Lyle WEISS
• 金额: $ 39.14万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-12 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10163221
• 关键词: Affect; Affinity; Alleles; Amino Acid Motifs; Animals; Architecture; Binding; Biochemical; Biological Assay; Cell Line; Cell Proliferation; Cell division; Cells; Cellular Morphology; Complex; Crystallization; Cytokinesis; Daughter; Dependence; Docking; Elements; Enzymes; Eukaryotic Cell; Evaluation; Event; Excision; Failure; Family; Genome Stability; Goals; Growth; Human; Human Cell Line; Knowledge; Ligands; Mammalian Cell; Maps; Mechanics; Methods; Molecular Genetics; Morphogenesis; Morphology; Output; Pathway interactions; Peptides; Phage Display; Phosphorylation; Phosphotransferases; Process; Property; Protein Family; Protein Kinase; Proteins; Proteome; Regulation; Reporting; Research; Research Design; Saccharomycetales; Signal Pathway; Signal Transduction; Site; Specificity; Staging; Structure; Substrate Interaction; System; Time; Tissues; Work; Yeasts; analog; carcinogenesis; daughter cell; gain of function; genetic analysis; in vivo; interest; novel; protein aminoacid sequence; protein transport; trafficking

PROJECT SUMMARY Eukaryotic appropriately commence daughter carcinogenesis processes cytokinesis cell division requires dependency relationships in which late events start only after early ones are completed. During cytokinesis, for example, distinct large-scale morphological processes in precise and consistent order to accomplish architectural reorganization that produces two cells. Failure of mechanical division of cells can drastically affect genome stability, contributing to and other important human maladies. mechanisms that coordinate cytokinetic are incompletely understood, and a subject of intense research interest. Notably, machinery of and regulators currently known to control it are highly conserved. Working in budding yeast, we Regulatory have discovered a system that blocks late events of cytokinesis when early ones are delayed or defective, allowing time for corrective mechanisms to function. This “checkpoint” pathway, termed “Enforcement of Cytokinesis Order” (ECO), works by stabilizing a multi-motif protein that blocks a cytokinesis-specific secretion function of the Ndr/Lats protein kinase Cbk1, a key component of a highly conserved “Ndr-hippo” signaling system. Hippo signaling pathways are crucial regulators of morphogenesis, proliferation, and differentiation of eukaryotic cells; Ndr/Lats kinases are their downstream-most signaling components. Our prior research contributed crucial analysis for understanding of these systems. We were was among the first to describe specific functions of a hippo signaling system in vivo, defined the distinctive Ndr kinase phosphorylation motif, reported the first canonical substrate docking by an AGC-family protein kinase, and contributed the first and only crystal structure of an Ndr/Lats kinase bound to a Mob co-activator. This project seeks to determine how the checkpoint mechanism we discovered blocks secretion of late cytokinesis proteins when early stages of the process are defective. It will include exploration of in vivo docking interactions with cytokinesis regulators, assessment of secretion organization during cytokinesis, and evaluation of the importance of Ndr-hippo regulation of key effector proteins of cytokinesis. We will also explore the hypothesis that human Ndr-hippo pathways participate in control of late cytokinesis, using two different approaches. In one, we will determine if human Ndr kinases engage in docking interactions with in vivo phosphorylation targets, using a highly optimized phage display approach to identify possible peptide motifs that interact with Ndr kinase – Mob co-activator complexes. We will use a new ligand footprinting method we developed to map peptide motif associations. In another approach, we will determine if human Ndr-hippo signaling functions in cytokinesis by constructing cell lines expressing only analog sensitive Ndr kinases. This will allow rapid and reversible inhibition of these kinases in cells undergoing both normal and defective cytokinesis.

项目摘要 真核生物 适当 开始 女儿 致癌作用 过程 细胞因子 细胞分裂需要依赖关系，其中较晚事件仅在早期开始之后开始 完全的。例如，在细胞因子中，不同的大规模形态学过程 精确，一致地完成建筑重组，产生两个 细胞。机械分裂细胞的失败会严重影响基因组稳定性，从而有助于 和其他重要的人类疾病。协调细胞力学的机制 不完全理解，也是一个强烈的研究兴趣的主题。值得注意的是，机械 目前已知可以控制它的监管机构是高度保守的。在萌芽的酵母中工作，我们 监管 已经发现了一个系统，该系统会阻止早期延迟或有缺陷时细胞因子的晚期事件， 允许时间正确的机制运行。这种“检查点”途径，称为“执行 细胞因子秩序（ECO），通过稳定阻断细胞因子特异性分泌的多氨基蛋白的作用 NDR/LATS蛋白激酶CBK1的功能，这是高度组成的“ NDR-HIPPO”信号的关键组成部分 系统。 河马信号通路是形态发生，增殖和分化的关键调节因子 真核细胞； NDR/LATS激酶是它们的下游信号传导组件。我们先前的研究 为了解这些系统的理解做出了重要分析。我们是第一个描述的人 河马信号系统在体内的特定功能定义了独特的NDR激酶磷酸化基序， 报道了AGC家庭蛋白激酶对接的第一个规范基板，并贡献了第一个和第一个和 仅与MOB共激活剂结合的NDR/LATS激酶的晶体结构。 该项目旨在确定我们发现最近的检查点机制如何分泌 当该过程的早期阶段有缺陷时，细胞因子蛋白。它将包括对体内的探索 与细胞因子调节剂相互作用，在细胞动力学期间的分泌组织评估以及 评估NDR-HIPPO调节细胞因子的关键效应蛋白的重要性。我们还将探索 人类NDR-Hippo途径参与了晚期细胞球运动的假设，使用两种不同 方法。在一个中，我们将确定人类NDR激酶是否参与与体内的互动 磷酸化靶标，使用高度优化的噬菌体显示方法来识别可能的肽基序 与NDR激酶 - MOB共激活因子复合物相互作用。我们将使用一种新的配体足迹方法 开发以绘制肽基序关联。在另一种方法中，我们将确定人类NDR-Hippo是否 信号传导通过仅表达模拟敏感NDR激酶的细胞系来在细胞因子中起作用。这 将允许在正常和缺陷的细胞中快速，可逆的这些激酶 细胞因子。