p120-catenin sub-family functions

描述（由申请人提供）：连环蛋白有助于所有动物细胞和组织的生物学，在不同的亚细胞区室中显示基本的胚胎和成体功能。例如，除了结合钙粘蛋白跨膜细胞-细胞粘附分子的内部（胞质）结构域之外，众所周知β-连环蛋白在Wnt发育信号传导中起重要作用，其中在细胞核内，它调节来自特定基因靶的转录。这些基因靶点不仅对正常胚胎发生很重要，而且对人类疾病包括结肠癌和黑色素瘤的进展也很重要。在这个更新的应用程序中，我们专注于较少了解的成员的连环蛋白家族的蛋白质，特别是成员的p120-连环蛋白亚家族（p120-连环蛋白; ARVCF-连环蛋白; δ-连环蛋白）的作用。一个主要的兴趣是上游Wnt通路信号如何产生一个网络的p120亚家族的反应，除了那些更好地了解β-连环蛋白。我们的目标包括阐明Wnt信号作用于p120亚家族成员的生化机制。对p120-catenin本身收集的证据表明，这种调节涉及激酶活性和p120-catenin蛋白稳定性的调节。与ARVCF和δ-连环蛋白相关的初步证据同样为Wnt途径反应性提供了可能性，例如，鉴定了两种涉及已知Wnt途径组分的新型δ-连环蛋白蛋白复合物。总的来说，我们的工作旨在扩大我们对连环蛋白生物学的理解范围，特别是关于p120-subF蛋白在Wnt信号传导中的新作用。在我们的研究中，我们将主要使用非洲爪蟾（青蛙）胚胎。该脊椎动物系统提供了外源构建体/试剂的容易引入（显微注射）、快速外部开发和许多生物化学和分子优势。对于功能分析，我们将利用敲低（吗啉代寡核苷酸），外源表达和拯救方法，结合发育表型的研究和蛋白质复合物形成或靶基因活性的生化和分子检测。为了补充我们在开发两栖动物胚胎方面的工作，我们将采用选择的哺乳动物细胞系，例如Wnt响应性HEK 293 T细胞。项目说明：Wnt信号通路是所有动物发育所必需的，而其适当的后期调节对于维持健康仍然很重要。例如，>90%的人结肠癌和黑色素瘤的进展涉及Wnt途径的病理激活。我们的建议检查一组属于p120-catenin亚家族的蛋白质的功能，我们有证据并假设参与Wnt信号传导，其研究将有助于了解正常发育和人类疾病。

DESCRIPTION (provided by applicant): The catenin proteins contribute to the biology of all animal cells and tissues, displaying essential embryonic and adult functions within differing sub-cellular compartments. For example, in addition to binding the inner (cytoplasmic) domain of cadherin trans-membrane cell-cell adhesion molecules, beta-catenin is well known to play an essential role in Wnt developmental signaling where within the nucleus it regulates transcription from specific gene targets. These gene targets are not only important for normal embryogenesis but also for the progression of human diseases including colon carcinoma and melanoma. In this renewal application, we focus upon the roles of less understood members of the catenin family of proteins, specifically members of the p120-catenin sub-family (p120- catenin; ARVCF-catenin; delta-catenin). A major interest is how upstream Wnt- pathway signals produce a network of p120 sub-family responses in addition to those better known for beta-catenin. Our aims include elucidating the biochemical mechanisms by which Wnt signals act upon p120 sub-family members. Evidence gathered for p120-catenin itself suggests that such regulation involves kinase activity and modulation of p120-catenin protein stability. Preliminary evidence relating to ARVCF and delta-catenin likewise leaves open the possibility of Wnt-pathway responsiveness, with for example, identification of two novel delta-catenin protein complexes involving known Wnt- pathway components. Overall, our work is aimed at expanding the scope of our understanding of catenin biology, with particular regard to the novel roles of p120-subF proteins in Wnt signaling. For our studies, we will predominantly employ Xenopus laevis (frog) embryos. This vertebrate system offers facile introduction (micro-injection) of exogenous constructs/ reagents, rapid external development and many biochemical and molecular advantages. For functional analysis, we will utilize knock-down (morpholino oligonucleotide), exogenous expression and rescue approaches in conjunction with the study of developmental phenotypes and biochemical and molecular tests of protein complex formation or target gene activity. To complement our work in developing amphibian embryos, we will employ select mammalian cell lines, for example, Wnt-responsive HEK293T cells. Project narrative: The Wnt signaling pathway is required for the development of all animals, while it's proper later regulation remains important in maintaining health. For example, the progression of >90% of human colon carcinomas and melanomas involves pathological activation of the Wnt pathway. Our proposal examines the functions of a group of proteins belonging to the p120-catenin sub-family, that we have evidence and hypothesize participates in Wnt signaling, and whose study will assist in understanding of both normal development and human disease.