PLAKOGLOBIN ORTHOLOGUES AND THE CELL

描述(摘自申请者的摘要)：我们目前的假设是 对Wnt信号的特异性反应由蛋白和蛋白共同决定 β-连环蛋白在某种程度上取决于它们的其他蛋白质结合和 修改。初步结果表明：(1)转基因表达 白蛋白抑制毛发生长；(2)β-连环蛋白的转基因表达 在乳腺中产生肿瘤；(3)白蛋白在 推测为CSK-3β磷酸化位点。我们的目标是解决以下问题 问题：(1B)白蛋白的作用是拮抗的、协同的还是 与β-连环蛋白互换以调节正常细胞中的细胞生长？ 将建立K14-deltaN89β-连环蛋白小鼠并比较它们的毛发表型 现有的K14-deltaN80-白蛋白小鼠。自发性肿瘤发病率 现有的MMTV-deltaN89β-连环蛋白和MMTV-deltaN80-Plakhemin小鼠将 被比较。拮抗或协同作用将通过这些杂交在体内进行测试 对小鼠和体外培养的转基因角质形成细胞进行检测 表达一个转基因对转基因细胞的定位、稳定性和稳定性的影响 另一种转基因产物的相互作用。(1B)是否上调监管 白蛋白激活FGFR？K14-deltaN80-白蛋白小鼠将被培育成 FGF5-/-安哥拉小鼠寻找长毛表型的中和作用 小路相交的证据。K14-DeltaN80-转基因白蛋白 角质形成细胞将被检查钙粘附素的上调和 上调FGFs作为潜在的FGFR配体。(2)白蛋白和 β-连环蛋白上调使表皮更容易发生肿瘤？一组 将对化学诱导的皮肤肿瘤进行筛查，以确定上调表达和 白蛋白和β-连环蛋白的重新定位和突变。K14转基因小鼠 将被测试以增强或抑制对肿瘤诱导的敏感性 (A)选育表达v-Rasha的小鼠(B0转化转基因 角质形成细胞与v-Rasha和移植到裸鼠；(C)标准化学 致癌方案。(3)白蛋白的糖基化是否在 蛋白质稳定性、连接形成和Wnt信号？亚细胞组分 角质形成细胞+/-连接形成和PC12细胞+/-WNT表达 检查是否有白蛋白糖浆的存在和程度。

DESCRIPTION (Adapted from applicant's abstract): Our current hypothesis is that specific responses to Wnt signals are determined by both plakoglobin and beta-catenin in a manner dependent on their other protein associations and modifications. Preliminary results show that (1) transgenic expression of plakoglobin inhibits hair growth; (2) transgenic expression of beta-catenin produces tumors in mammary gland; (3) plakoglobin is glycosylated in the putative CSK-3beta phosphorylation site. Our aims address the following questions: (1b) does plakoglobin act antagonistically, synergistically or interchangably with beta-catenin to regulate cell growth in normal cells? K14-deltaN89beta-catenin mice will be created and their hair phenotype compared to the existing K14-deltaN80-plakoglobin mice. The spontaneous tumor incidence of existing MMTV-deltaN89beta-catenin and MMTV-deltaN80-plakoglobin mice will be compared. Antagonism or synergy will be tested in vivo by crossing of these pairs of mice and in vitro by examining cultured transgenic keratinocytes for the effects of expressing one transgene on the localization, stability and interaction of the produce of the other transgene. (1b) does up-regulated plakoglobin activate FGFR? K14-deltaN80-plakoglobin mice will be bred to FGF5-/- angora mice to look for counteraction of the long hair phenotype as evidence of intersection of the pathways. K14-deltaN80-plakoglobin trangenic keratinocytes will be examined for up-regulation of cadherins and for up-regulation of FGFs as potential FGFR ligands. (2) Does plakoglobin and beta-catenin up-regulation predispose epidermis to tumorigenesis? A panel of chemically induced skin tumors will be screened for up-regulation and relocation and mutations of plakoglobin and beta-catenin. K14 transgenic mice will be tested for enhanced or suppressed susceptibility to tumor induction by (a) breeding to mice expressing v-rasHa: (b0 transforming transgenic keratinocytes with v-rasHa and grafting to athymic mice; (c) standard chemical carcinogenesis protocols. (3) does glycosylation of plakoglobin play a role in protein stability, junction formation and Wnt-signaling? subcellular fractions of keratinocytes+/- junction formation and PC12 cells +/- wnt expression will be examined for presence and extent of plakoglobin glycosylaiton.