Vangl2与Scrib1相互作用以及vangl2的酵母双杂交筛选胎脑库研究

发布时间：2018-01-19 02:38

  本文关键词： Vangl2 Scrib1 WNT NTDs PTPN18 酵母双杂交 信号转导　出处：《兰州大学》2006年硕士论文　论文类型：学位论文

【摘要】：Vangl2为高度保守的膜蛋白,它是一个521个氨基酸的跨膜蛋白,N-端有四个跨膜结构域,C-端在细胞质面,含有一个PDZ结合结构域,与细胞内信号转导或与细胞内其他信号分子相互作用有关。Vangl2是一个非经典WNT信号通路的一种成分,与细胞平面极化(PCR,planar cell polarity)相关,调节细胞排布的方向,Vangl2突变导致小鼠产生looped-tail表型,这种小鼠有神经管闭合缺陷。 Scrib1为一种抑癌基因,文献报道,用果蝇胚胎上皮细胞、卵泡上皮细胞、视盘上皮细胞为研究对象,发现Scrib1突变导致上皮组织极化失常,失去上皮细胞的正常结构,使其过度生长。Scrib1插入突变使小鼠产生Circletail(Crc)表型,与Vangl2突变一样,这种小鼠也有神经管闭合缺陷,同时越来越多的文献报道Scrib1也参与PCP信号过程,与Scrib1相关的分子也与Vangl2存在直接或者间接的相互联系,提示它们在信号通路上存在交叉。 本文主要研究Vangl2与Scrib1的相互作用,用pull-down证明Scrib1和Vangl2存在相互作用,并采用了酵母双杂交技术研究Vangl2的信号途径,我们选择构建编码Vangl2的C-末端的基因片段为酵母双杂交的诱饵(bait),筛选胎脑库得到了一些候选基因,其中一个为非受体类型蛋白酪氨酸磷酸酯酶(protein tyrosine phosphatase non-receptor)PTPN18。酪氨酸激酶对原癌基因的激活在癌细胞信号转导中是很普遍的一种现象,酪氨酸磷酸化也由蛋白酪氨酸磷酸酯酶/酪氨酸激酶控制,一些蛋白酪氨酸磷酸酯酶含有SHP2结构域,能正向调节生长因子受体的信号通路,具有致癌性。 在WNT信号通路中:WNT活化Fz信号,Fz磷酸化Dsh,Dsh抑制GSK-3β,使β-catenin累积,调节各种基因转录,β-catenin的磷酸化和去磷酸化对于信号通路有着重要的作用。我们注意到Vangl2与Fz受体一样也是一个膜蛋白,而它与PTPN18这个酪氨酸磷酸酯酶存在相互作用,同时本实验室以前用酵母双杂证明Scrib1和proline-serine-threonine phosphatase interacting protein 1(pstpip 1)蛋白相互作用,而通过www.bind.ac数据库,查到pstpip1和ptpn18有相互作用,我们有理由认为这个Vangl2膜受体受到细胞外分子信号的刺激后,至少形成Vangl2-ptpn18-pstpip1-Scrib1的复合体,使某个下游分子或者蛋白复合体中的某个成分磷酸/去磷酸化,导致一系列的效应。
[Abstract]:Vangl2 is a highly conserved membrane protein. It is a 521 amino acid transmembrane protein with four transmembrane domains C- terminal in the cytoplasm and a PDZ binding domain. Vangl2 is a component of a nonclassical WNT signaling pathway associated with intracellular signal transduction or interaction with other intracellular signaling molecules. Planar cell polarity). The mutation of Vangl2, which regulates the distribution of cells, leads to the production of looped-tail phenotype in mice. This mouse has a neural tube closure defect. Scrib1 is a tumor suppressor gene. The embryonic epithelial cells, follicular epithelial cells and optic disk epithelial cells of Drosophila melanogaster were used in this study. It was found that Scrib1 mutation resulted in abnormal polarization of epithelial tissue and loss of normal structure of epithelial cells. The insertion mutation of Scrib1 caused the mice to produce Circletaila Crcc phenotype, which, like the Vangl2 mutation, had neural tube closure defects. At the same time, more and more literatures have reported that Scrib1 is also involved in the PCP signal process, and Scrib1 related molecules are directly or indirectly related to Vangl2. These results suggest that they are intersected in signal pathway. In this paper, the interaction between Vangl2 and Scrib1 is studied, and the existence of interaction between Scrib1 and Vangl2 is proved by pull-down. Yeast two-hybrid technique was used to study the signal pathway of Vangl2. We selected the C-terminal gene fragment encoding Vangl2 as the bait of yeast two-hybrid. A number of candidate genes were obtained from the screening of fetal brain banks, one of which was a non-receptor type protein tyrosine phosphatase protein. Tyrosine phosphatase non-receptor). PTPN18. the activation of proto-oncogene by tyrosine kinase is a common phenomenon in cancer cell signal transduction. Tyrosine phosphorylation is also controlled by protein tyrosine phosphatase / tyrosine kinase. Some protein tyrosine phosphatase contain SHP2 domain and can positively regulate the signal pathway of growth factor receptor. Carcinogenic. In the WNT signaling pathway, WNT activates FZ phosphorylation of GSK-3 尾, inhibits the accumulation of 尾 -catenin, and regulates the transcription of various genes. The phosphorylation and dephosphorylation of 尾 -catenin play an important role in the signaling pathway. We note that Vangl2 is a membrane protein as well as FZ receptor. And it interacts with PTPN18, a tyrosine phosphatase. At the same time, we used yeast double hybrid to prove Scrib1 and proline-serine-threonine phosphatase. Interacting protein 1. Pstpip 1) protein interaction. Through the www.bind.ac database, the interaction between pstpip1 and ptpn18 was found. It is reasonable to believe that this Vangl2 membrane receptor is stimulated by extracellular molecular signals and at least forms a complex of Vangl2-ptpn18-pstpip1-Scrib1. Phosphorylation of a component of a downstream molecule or protein complex leads to a series of effects.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R341;Q78

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