Ubiquitin-like protein modifications in planar cell polarity

平面细胞极性中的泛素样蛋白修饰

基本信息

批准号：
9240642
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
项目类别：
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/9240642
关键词：
APC2 gene Address Adult 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 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 Mosaicism 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 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 experimental study follow-up gastrulation genetic approach genetic regulatory protein genome-wide analysis in vivo insight intercalation loss of function member negative affect novel planar cell polarity polarized cell protein degradation public health relevance receptor skin organogenesis ubiquitin ligase

项目摘要

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：上皮细胞的极化在两个轴上是明显的，在普遍存在的顶端-基底外侧轴和作为第二轴的上皮平面内，后者通常被称为平面细胞极性（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、毛囊方向或内脏器官的细胞排列，如内耳上皮及其感觉纤毛。在果蝇中，所有成年角质层结构和器官都显示显着的 PCP 特征，使得果蝇 PCP 建立的研究成为发育模式和一般疾病过程的范例。对果蝇的分析已经建立了一条围绕卷曲 (Fz)/PCP 核心因子盒和相关调节因子的保守分子途径。这一核心 Fz/PCP 通路及其调节成分在整个进化过程中都是保守的，不仅在上皮器官中，而且在哺乳动物的定向细胞迁移中调节细胞极化的许多方面原肠胚形成和神经形成过程中间充质细胞的形成。尽管核心 Fz/PCP 因子之间相互作用的框架正在出现，但人们对它们相互作用背后的实际分子机制知之甚少。该应用的范围是跟踪从全基因组筛选中非常有趣的基因鉴定，寻找核心 PCP 因子的新型调节剂。基于有趣的初步数据，我们提出的具体目标是 (1) 解决后期促进复合物 (APC/C) 泛素连接酶活性的作用，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)定义由 CG15283 编码的新型泛素样修饰剂在 PCP 建立中的功能，以及它如何在 PCP 信号传导过程中调节核心 Fz/PCP 因子或 APC/C 成分的活性，以及(3) 识别APC/C的PCP特异性底物并定义APC/C对它们的分子调控。我们已经建立了几种测定方法，使我们能够通过果蝇体内功能研究、生化实验和细胞培养分析的结合来实现这些目标。由于细胞周期控制之外的 APC/C 的作用和保守的 CG15283 肽的功能分别在很大程度上模糊或完全未知，我们的应用将提供对这些机制的初步见解。 Fz/PCP 的建立与多种医学异常有关，包括耳聋、癌症、多囊肾病和纤毛病。因此，在此应用中获得的信息不仅将增进我们对 PCP 调节和器官模式的理解，而且还将在多种疾病背景下具有医学相关性。