Ubiquitin-like protein modifications in planar cell polarity

平面细胞极性中的泛素样蛋白修饰

• 批准号: 8628229
• 负责人: Marek Mlodzik
• 金额: $ 32.21万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8628229
• 关键词: APC2 gene; Address; Adult; Affect; Anaphase; Apical; Architecture; Biochemical; Biological Assay; Caenorhabditis elegans; Cell Culture Techniques; Cell Cycle Progression; Cell Cycle Proteins; Cell Cycle Regulation; Cell Polarity; Cells; Cilia; Complex; Data; Development; Disease; Drosophila genus; Employee Strikes; Epidermis; Epithelial; Epithelial Cells; Epithelium; Evolution; Genes; Genetic; Hair; Hair follicle structure; Human; Insecta; Invertebrates; Labyrinth; Ligase; Link; Maintenance; Malignant Neoplasms; Mammalian Oviducts; Mammals; Mediating; Mediator of activation protein; Medical; Mesenchymal; Mitotic; Molecular; Morphogenesis; Mutation; Names; Organ; Organogenesis; Pathway interactions; Pattern; Peptides; Physiological; Polycystic Kidney Diseases; Post-Translational Protein Processing; Process; Proteins; Proto-Oncogenes; Regulation; Role; Sensory; Signal Transduction; Skin; Structure; System; Tissues; Tumor Suppressor Proteins; Ubiquitin; Ubiquitin Like Proteins; Ubiquitination; Vertebrates; Whole Organism; anaphase-promoting complex; base; carcinogenesis; cell motility; cell type; ciliopathy; deafness; follow-up; gastrulation; genetic regulatory protein; genome wide association study; in vivo; insight; intercalation; interest; loss of function; member; novel; protein degradation; public health relevance; receptor; research study; ubiquitin ligase

Polarization of epithelial cells is evident in two axes, in the ubiquitous apical-basolateral axis and within the plane of the epithelium as a second axis, the latter generally referred to as Planar Cell Polarity (or PCP). Examples of PCP are present in almost all organs, and in mammals, for example, most obvious in aspects of skin development with hair follicle orientation or cellular arrangements in internal organs, like the inner ear epithelium with its sensory cilia. In Drosophila, all adult cuticular structures and organs display striking PCP features, which makes the study of PCP establishment in Drosophila serve as a paradigm for the process in developmental patterning and disease in general. Analyses in Drosophila have established a conserved molecular pathway anchored around the Frizzled (Fz)/PCP core factor cassette and associated regulatory factors. This core Fz/PCP pathway and its regulatory components are conserved throughout evolution regulating many aspects of cellular polarization not only in epithelial organs, but in mammals also in directed cell migration of mesenchymal cells during gastrulation and neurulation. Although a framework of the interactions among the core Fz/PCP factors is emerging, little is known about the actual molecular mechanisms underlying their interactions. The scope of this application is to follow-up on very interesting gene identifications from a genome wide screen for novel regulators of the core PCP factors. Based on interesting preliminary data, we propose as Specific Aims to (1) address the role of the Ubiquitin-ligase activity of the Anaphase Promoting Complex (APC/C) and define which components of this complex act in PCP and how, (2) define the function of the novel ubiquitin-like modifier encoded by CG15283 in PCP establishment and how it might regulate the activity of core Fz/PCP factors or APC/C components during PCP signaling, and (3) identify the PCP specific substrates of the APC/C and define their molecular regulation by the APC/C. We have established several assays that will allow us to address these aims via a combination of functional in vivo studies in Drosophila, biochemical experiments, and cell culture analyses. As the roles of the APC/C outside cell cycle control and a function of the conserved CG15283 peptide are largely obscure or completely unknown, respectively, our application will provide first insight(s) into the mechanism(s) of these. Fz/PCP establishment has been linked to several medical abnormalities, including deafness, cancer, polycystic kidney disease, and ciliopathies. Thus the information acquired in this application will both advance our understanding of PCP regulation and organ patterning, and will also be of medical relevance in several disease contexts.

上皮细胞的极化在两个轴上很明显，在普遍存在的顶端-基底外侧轴和内 上皮平面作为第二轴，后者通常称为平面细胞极性（或 PCP）。 五氯苯酚的例子几乎存在于所有器官中，例如在哺乳动物中，最明显的是 皮肤发育与毛囊方向或内脏器官（如内耳）的细胞排列有关 上皮及其感觉纤毛。在果蝇中，所有成年角质层结构和器官都表现出惊人的 PCP 特点，这使得果蝇 PCP 建立的研究可以成为果蝇过程的范例。 一般发育模式和疾病。果蝇分析建立了一个保守的 锚定在卷曲 (Fz)/PCP 核心因子盒和相关监管周围的分子途径 因素。这一核心 Fz/PCP 途径及其调控组件在整个进化过程中得到保留 不仅在上皮器官中，而且在哺乳动物中也以定向方式调节细胞极化的许多方面 原肠胚形成和神经形成期间间充质细胞的细胞迁移。虽然一个框架 核心 Fz/PCP 因子之间的相互作用正在出现，但人们对实际的分子机制知之甚少。 它们相互作用的机制。这个应用的范围是追踪非常有趣的基因 从全基因组筛选中鉴定出核心 PCP 因子的新型调节剂。基于有趣的 初步数据显示，我们提出的具体目标是 (1) 解决泛素连接酶活性的作用 后期促进复合物 (APC/C) 并定义该复合物的哪些成分在 PCP 中起作用以及如何起作用，(2) 定义 CG15283 编码的新型泛素样修饰剂在 PCP 建立中的功能及其如何实现 可能在 PCP 信号传导过程中调节核心 Fz/PCP 因子或 APC/C 成分的活性，并且 (3) 识别 APC/C 的 PCP 特异性底物并定义 APC/C 对其的分子调节。我们有 建立了几种测定方法，使我们能够通过体内功能的组合来解决这些目标 果蝇研究、生化实验和细胞培养分析。作为APC/C以外的角色 细胞周期控制和保守的 CG15283 肽的功能很大程度上不清楚或完全不清楚 分别未知，我们的应用程序将提供对这些机制的初步见解。氟/五氯苯酚 建立与多种医学异常有关，包括耳聋、癌症、多囊肾 疾病和纤毛病。因此，在此应用程序中获得的信息将增进我们的理解 PCP 监管和器官模式，并且在多种疾病背景下也具有医学相关性。