Dsh/Dvl Phosphorylation and Signaling Outcome

Dsh/Dvl 磷酸化和信号转导结果

• 批准号: 7714941
• 负责人: Marek Mlodzik
• 金额: $ 25.43万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-15 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7714941
• 关键词: Address; Animal Model; Animal Testing; Apoptosis; Biochemical; Biological Assay; Cell Culture Techniques; Cell Polarity; Cell physiology; Cells; Complement; Data; Defect; Development; Diagnostic; Disease; Drosophila genus; Embryo; Epithelium; Event; Excision; Eye; FZD2 gene; Family; Felis catus; Generations; Growth Factor; Hand; Homeostasis; Immune Sera; Invertebrates; Link; Malignant Neoplasms; Mammalian Cell; Mass Spectrum Analysis; Mediating; Medical; Membrane; Molecular; Outcome; Pathway interactions; Pattern; Phospho-Specific Antibodies; Phosphorylation; Phosphotransferases; Photoreceptors; Physiological; Polycystic Kidney Diseases; Protein Tyrosine Kinase; Proto-Oncogenes; Recruitment Activity; Regulation; Research; Role; Sampling; Signal Pathway; Signal Transduction; Site; Specificity; Staging; Surveys; Time; Tissues; Transducers; Tumor Suppressor Proteins; Tyrosine Phosphorylation; Visual Fields; Wing; angiogenesis; base; cellular development; fly; in vitro Assay; in vivo; mutant; novel; public health relevance; receptor; research study; response; tool

DESCRIPTION (provided by applicant): Frizzled (Fz) receptors act as signal transducers of Wnt growth factors via several effector pathways, notably Wnt/2-catenin (2-cat) and Fz/planar cell polarity (PCP) signaling. Fz/PCP and Wnt-Fz/2-cat signaling diverge downstream of the cytoplasmic factor Dishevelled (Dsh). Often both pathways act in the same tissues, and a tight regulation of signaling specificity is essential for normal development and cellular homeostasis. In deregulated scenarios, selection of the wrong Fz-pathway often leads to disease (i.e. PKD or cancer). Although a molecular framework for both pathways is established, very little is known about the regulation of Wnt/Fz-Dsh signaling specificity. This application investigates the molecular aspects of signaling specificity distinguishing between Wnt-Fz/2-Cat and Fz/PCP signaling. We will address this in Drosophila (the model organism where both pathways were discovered) and in mammalian cell culture. The specific focus is the regulation of Dsh by phosphorylation. Dsh is a cytoplasmic factor shared between both pathways and recruited to the membrane by Fz receptors. Based on preliminary data we hypothesize that Dsh is differentially phosphorylated in a pathway specific manner. A combination of cell culture, in vivo, and biochemical studies will dissect the phosphorylation events on Dsh that switch its function between the Wnt- Fz pathways. The Specific Aims are: 1. To determine specific phosphorylation event(s) on Dsh and their role in the selection of the signaling outcome; and 2. To identify and functionally define Tyrosine kinases acting in Dsh regulation and pathway specific responses. Our mass-spectrometry studies indicated that differential phosphorylation of Dsh contributes to pathway selection and we have identified several kinases acting on Dsh. Physiological and medical relevance of the phosphorylation events will be established in vivo. This will be complemented by the generation of specific antisera to the relevant phosphorylated sites in Dsh. These are likely to have diagnostic potential. PUBLIC HEALTH RELEVANCE: The regulation and selection of the correct Wnt signaling pathways is an essential feature of development and tissue function and homeostasis. Deregulation of Wnt is linked to many diseases, ranging from ciliopathies and angiogenesis defects, to cancer (e.g. several pathway components are tumor suppressors or proto-oncogenes). This application addresses the molecular regulation of Dishvelled a key component in Wnt/Frizzled-signaling specificity; the information acquired here will be of high significance for several medical disorders.

描述（由申请人提供）：卷曲（Fz）受体通过多种效应途径，特别是Wnt/2-连环蛋白（2-cat）和Fz/平面细胞极性（PCP）信号传导，充当Wnt生长因子的信号转导器。 Fz/PCP 和 Wnt-Fz/2-cat 信号传导在细胞质因子 Disheveled (Dsh) 下游出现分歧。通常，这两种途径都在同一组织中起作用，信号特异性的严格调节对于正常发育和细胞稳态至关重要。在放松管制的情况下，选择错误的 Fz 通路通常会导致疾病（即 PKD 或癌症）。尽管两条途径的分子框架都已建立，但对 Wnt/Fz-Dsh 信号传导特异性的调节知之甚少。此应用研究了区分 Wnt-Fz/2-Cat 和 Fz/PCP 信号传导的信号传导特异性的分子方面。我们将在果蝇（发现这两种途径的模型生物）和哺乳动物细胞培养物中解决这个问题。具体重点是磷酸化对 Dsh 的调节。 Dsh 是两种途径之间共享的细胞质因子，并由 Fz 受体募集到细胞膜上。根据初步数据，我们假设 Dsh 以通路特异性方式差异磷酸化。细胞培养、体内和生化研究的结合将剖析 Dsh 上的磷酸化事件，这些事件在 Wnt-Fz 途径之间切换其功能。具体目标是： 1. 确定 Dsh 上的特定磷酸化事件及其在信号转导结果选择中的作用； 2. 鉴定并在功能上定义在 Dsh 调节和通路特异性反应中发挥作用的酪氨酸激酶。我们的质谱研究表明 Dsh 的差异磷酸化有助于途径选择，并且我们已经鉴定了几种作用于 Dsh 的激酶。将在体内建立磷酸化事件的生理学和医学相关性。这将通过针对 Dsh 中相关磷酸化位点生成特异性抗血清来补充。这些可能具有诊断潜力。公共健康相关性：正确 Wnt 信号通路的调节和选择是发育、组织功能和体内平衡的基本特征。 Wnt 失调与许多疾病有关，从纤毛病和血管生成缺陷到癌症（例如，几个途径成分是肿瘤抑制基因或原癌基因）。该应用解决了 Dishvelled 的分子调控问题，Dishvelled 是 Wnt/Frizzled 信号特异性的关键成分；这里获得的信息对于多种医学疾病具有重要意义。