Tyrosine Phosphorylation of LRP6 and its role in regulation Wnt signaling

LRP6 酪氨酸磷酸化及其在 Wnt 信号传导中的调节作用

• 批准号: 214904911
• 负责人: Dr. Gary Davidson
• 金额: --
• 依托单位: Institut für Toxikologie und Genetik (ITG) (aufgelöst)
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2012
• 资助国家: 德国
• 起止时间: 2011-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/214904911?language=en
• 关键词: Tyrosine; Phosphorylation; LRP6; its; role

Metazoans use Wnt/β-catenin signaling to control critical aspects of cell behavior and deregulated Wnt signaling is strongly implicated in human disease. We perform cell culturebased expression screening to identify new modifiers that effect specific Wnt pathway components. A key focus of our work is to understand how cells control reception of Wnt signaling via low-density lipoprotein receptor-related-protein 6 (LRP6). Although serine and threonine phosphorylation has been well studied, the potential role of tyrosine phosphorylation of LRP6 has not been investigated. Using pan phospho-specific antibodies we screened a kinase cDNA library by SDS-PAGE / Immunoblot analysis and identified a novel LRP6 tyrosine kinase. Preliminary work demonstrates that it specifically interacts with LRP6, inhibits LRP6/Wnt signaling and is itself regulated by Wnt signaling. These finding indicate that tyrosine phosphorylation of LRP6 plays an important role in regulating Wnt signaling. Here we propose to analyze in detail how this kinase functions to regulate LRP6, using molecular biology, cell biology as well as in-vivo experiments. Biochemical experiments will be performed using cell culture and physiological relevance will be addressed using both zebrafish and Xenopus.

后生动物使用Wnt/β-连环蛋白信号来控制细胞行为的关键方面，Wnt信号的失调与人类疾病密切相关。我们进行基于细胞培养的表达筛选，以确定影响特定Wnt通路成分的新修饰剂。我们工作的一个重点是了解细胞如何通过低密度脂蛋白受体相关蛋白6 （LRP6）控制Wnt信号的接收。虽然丝氨酸和苏氨酸磷酸化已经得到了很好的研究，但酪氨酸磷酸化LRP6的潜在作用尚未得到研究。利用泛磷特异性抗体，我们通过SDS-PAGE /免疫印迹分析筛选了一个激酶cDNA文库，并鉴定了一个新的LRP6酪氨酸激酶。初步研究表明，它与LRP6特异性相互作用，抑制LRP6/Wnt信号传导，并受Wnt信号传导调节。这些发现表明LRP6酪氨酸磷酸化在调节Wnt信号传导中起重要作用。本文拟从分子生物学、细胞生物学和体内实验等方面详细分析该激酶调控LRP6的作用机制。生化实验将使用细胞培养进行，生理相关性将使用斑马鱼和爪蟾进行。