Characterizing Novel Wnt Signaling Pathway Components

描述（由申请人提供）：在所有脊椎动物中，经典Wnt信号传导调节胚胎发育和细胞命运以及增殖决定。还有一种非经典信号通路调节平面细胞极性（PCP）。在这些信号通路中起作用的几种蛋白质的突变与许多上皮癌有关。初步工作表明，一种新的果蝇G蛋白偶联受体（GPCR）可能在Wnt信号转导中发挥作用。我们还发现了导致无翼重组表型的基因组损伤。我们建议，这种GPCR和这个身份不明的基因代表新的Wnt信号传导因子，我们将使用遗传和生物化学技术来确定它们的具体作用。我们将绘制基因组损伤，然后通过产生无效等位基因并测量其对下游信号传导事件的影响来定义其在Wnt通路中的作用。我们将使用免疫荧光和细胞生物学研究确定GPCR敲除的表型。然后，我们将使用遗传学将这种新基因置于Wnt途径中。然后将进行类似的细胞生物学和遗传实验，以确定GPCR在PCP信号传导中的作用。

DESCRIPTION (provided by applicant): Canonical Wnt signaling regulates embryonic development and cell fate and proliferation decisions in all vertebrates. There is also a non-canonical signaling pathway that regulates planar cell polarity (PCP). Mutations in several of the proteins acting in these signaling pathways are associated with numerous epithelial cancers. Preliminary work indicates that a novel Drosophila G-protein coupled receptor (GPCR) could have a role in Wnt signaling. We have also identified a genomic lesion resulting in a phenotype reassembling wingless. We propose that this GPCR and this unidentified gene represent novel Wnt signaling factors and we will determine their specific roles using genetic and biochemical techniques. We will map the genomic lesion and then define its role in the Wnt pathway by generating a null allele and measuring its effect on downstream signaling events. We will determine the phenotype of a GPCR knock out using immunofluorescence and cell biological studies. We will then use genetics to place this novel gene in the Wnt pathway. Similar cell biological and genetic experiments will then be done to determine the GPCR's role in PCP signaling.