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是显而易见的，在无处不在的根尖轴和内部的极化。 上皮的平面为第二轴，后者通常称为平面细胞极性（或PCP）。 PCP的示例几乎存在于所有器官中，例如，在哺乳动物中，最明显的是 皮肤发育具有毛囊取向或内部器官内部器官的细胞排列 上皮及其感觉纤毛。在果蝇中，所有成人表皮结构和器官都显示出醒目的PCP 功能，使果蝇的PCP建立研究是该过程的范式 发育模式和疾病总体上。果蝇的分析已经建立了保守的 分子途径锚定在卷曲的（FZ）/PCP核心因子盒和相关调节 因素。该核心FZ/PCP途径及其调节组件在整个演化过程中都保守 调节细胞极化的许多方面，不仅在上皮器官中，而且在哺乳动物中也有指导 间充质细胞的细胞迁移在胃和神经化过程中。虽然是 核心FZ/PCP因子之间的相互作用是出现的，对实际分子知之甚少 其相互作用的基础机制。此应用的范围是跟进非常有趣的基因 来自基因组宽屏幕的鉴定，用于核心PCP因素的新型调节剂。基于有趣的 初步数据，我们提出的是特定目的，目的是（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PCP中该复合物的哪些组成部分以及如何（2） 定义由CG15283在PCP机构中编码的新型泛素样修饰符的功能及其如何 可能调节核心FZ/PCP因子或PCP信号期间APC/C组件的活性，（3）确定 APC/C的PCP特异性底物，并通过APC/C定义其分子调节。我们有 建立了几种测定法，使我们能够通过体内功能的结合来解决这些目标 果蝇，生化实验和细胞培养分析的研究。作为APC/C外部的角色 细胞周期控制和保守CG15283肽的功能在很大程度上是晦涩或完全晦涩的 我们的应用程序分别将对这些应用程序的机制提供第一个洞察力。 FZ/PCP 建立与几种医学异常有关，包括耳聋，癌症，多囊性肾脏 疾病和纤毛病。因此，本应用程序中获得的信息既可以提高我们的理解 PCP调节和器官模式的范围，在几种疾病情况下也将具有医学相关性。