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One signal, two pathways: Analysis of how a single Wnt ligand can initiate discrete signaling pathways through the activation of two distinct receptors.

机译:一种信号,两种途径:分析单个Wnt配体如何通过两种不同受体的激活而引发离散的信号途径。

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摘要

Cell to cell communication is vital throughout the development of multicellular organisms and during adult homeostasis. One way in which cell communication occurs is through the secretion of signaling molecules that are received by neighboring responding cells. Wnt ligands comprise a large family of secreted hydrophobic glycoproteins that control a variety of developmental and adult processes in all metazoan organisms. As inappropriate Wnt signaling contributes to cancer and other degenerative disorders, much effort has been made to understand the ways in which Wnt signaling is activated and regulated. In the past, controlled in vitro studies of Wnt signal activation were hampered by a lack of active, soluble protein. However, purified Wnt proteins can now be generated and questions regarding how various Wnt proteins exert their pleiotropic effects can finally be studied.;In this thesis I will address the fundamental question of whether all Wnt family members signal in the same manner or whether different Wnt family members initiate discrete signaling pathways by engaging different receptors. In the most well understood "canonical" Wnt signaling pathway, Wnt binding to Frizzled receptors induces beta-Catenin protein stabilization and entry into the nucleus, where it complexes with TCF/LEF transcription factors to affect the transcription of target genes. Through the use of purified Wnt proteins, I will show that Wnt5a protein can both activate and inhibit canonical Wnt signaling in the presence of the appropriate receptor. Wnt5a-mediated inhibition of canonical Wnt signaling requires the receptor tyrosine kinase Ror2 and occurs at the level of gene transcription. Through mutational analysis, I will also show which regions of the mRor2 receptor are necessary for mediating the Wnt5a inhibitory signal. Lastly, to gain insight into the role Ror2 plays in inhibiting canonical Wnt signaling in vivo, I will explore the expression pattern of the Ror2 receptor using mono- and polyclonal antibodies that I generated for that purpose.
机译:在整个多细胞生物的发展过程中以及成年体内的稳态过程中,细胞间的通讯至关重要。细胞通讯发生的一种方式是通过分泌被相邻反应细胞接收的信号分子。 Wnt配体包含一个大家族的分泌性疏水糖蛋白,可控制所有后生生物中的各种发育和成年过程。由于不适当的Wnt信号传导会导致癌症和其他退行性疾病,因此人们做出了很多努力来了解Wnt信号活化和调控的方式。过去,缺乏活性的可溶蛋白阻碍了Wnt信号激活的体外对照研究。然而,现在可以生成纯化的Wnt蛋白,并且最终可以研究有关各种Wnt蛋白如何发挥其多效性作用的问题。家庭成员通过参与不同的受体来启动离散的信号传导途径。在最广为人知的“规范” Wnt信号传导途径中,Wnt与卷曲蛋白受体的结合可诱导β-连环蛋白稳定并进入细胞核,并与TCF / LEF转录因子复合,从而影响靶基因的转录。通过使用纯化的Wnt蛋白,我将证明Wnt5a蛋白在适当受体存在下既可以激活也可以抑制经典Wnt信号传导。 Wnt5a介导的经典Wnt信号传导抑制需要受体酪氨酸激酶Ror2并在基因转录水平上发生。通过突变分析,我还将显示mRor2受体的哪些区域对于介导Wnt5a抑制信号是必需的。最后,为了深入了解Ror2在体内抑制经典Wnt信号传导中的作用,我将使用为此目的而产生的单克隆和多克隆抗体探索Ror2受体的表达模式。

著录项

  • 作者

    Mikels, Amanda Jane.;

  • 作者单位

    Stanford University.;

  • 授予单位 Stanford University.;
  • 学科 Biology Molecular.
  • 学位 Ph.D.
  • 年度 2007
  • 页码 158 p.
  • 总页数 158
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分子遗传学;
  • 关键词

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