Regulation of Hippo pathway signaling by mechanical forces

项目摘要 这个项目的长期目标是揭示转录共激活子 而癌基因YAP受LATS和SRC激酶的调节，以响应机械刺激， 这可能会带来更好的癌症治疗方法，并改进干细胞疗法。航站楼 河马途径激酶LATS抑制YAP核定位，但也具有较差的特性 在胞质分裂中发挥重要作用，而SRC酪氨酸激酶促进YAP核 本地化。YAP的核定位受到多种刺激的调节，如肌动蛋白细胞骨架， 控制密度依赖抑制的底物硬度、细胞分离、细胞拥挤和拉伸 包括生长、组织修复、干细胞增殖和分化。当在核中时，YAP 促进细胞存活和增殖。在这里，我们将确定LATS是如何由这两个 细胞-细胞连接处的张力控制密度依赖的细胞生长抑制和有丝分裂期间 控制胞质分裂。鉴定特定的有丝分裂途径可能使LATS活性 在不干扰细胞分裂的情况下促进组织再生。我们还将测试一个 SRC激酶如何与AMOT蛋白协同反应细胞黏附的假说 和其他刺激物将YAP靶向细胞核。在具体目标1中，我们将确定几个 在细胞-细胞连接处组成张力传感器的蛋白质共同作用，调节Lats的活性。 对细胞张之间的细胞张力的反应。在特定目标2中，我们将检验一个假设 SRC将很多人从YAP的抑制者变成了激活者。理解这条途径将是 对于确定增加AMOT水平的药物是否是治疗AMOT的合适方法很重要 考虑到癌症。在特定的目标3中，我们将确定lats在有丝分裂中是如何被激活的，其身份 有丝分裂底物，以及它如何促进胞质分裂。总体而言，这项工作将揭示LATS和YAP如何 是受特定刺激调节的。这些研究将对我们理解 抑制肿瘤和组织再生，并可能导致更好的方法来操纵这些 重要的过程。

Project Summary The long term goal of this project is to uncover the pathways by which the transcriptional co-activator and oncogene YAP is regulated by the LATS and SRC kinases in response to mechanical stimuli, which could lead to better treatments for cancer, and improved stem cell therapies. The terminal Hippo pathway kinase LATS inhibits YAP nuclear localization, but also has a poorly characterized but important functions in cytokinesis, whereas the SRC tyrosine kinase promotes YAP nuclear localization. YAP nuclear localization is regulated by diverse stimuli such as the actin cytoskeleton, substrate stiffness, cell detachment, cell crowding, and stretch to control density dependent inhibition of growth, tissue repair and stem cell proliferation and differentiation. When in the nucleus, the YAP promotes cell survival and proliferation. Here we will determine how LATS is regulated by both tension at cell-cell junctions to control density dependent inhibition of cell growth and during mitosis to control cytokinesis. Identification of specific mitotic pathways may allow LATS activity to be manipulated to promote tissue regeneration without interfering with cell division. We will also test a hypothesis for how the SRC kinase collaborates with the AMOT protein in response to cell adhesion and other stimuli to target YAP to the nucleus. In Specific Aim 1, we will determine how several proteins that comprise a tension sensor at cell-cell junctions work together to regulate LATS activity in response to cellular tension across sheets of cells. In Specific Aim 2 we will test a hypothesis that SRC turns AMOT from an inhibitor of YAP into an activator. Understanding this pathway will be important for determining whether drugs that increase AMOT levels are appropriate therapies for a given cancer. In Specific Aim 3 we will determine how LATS is activated in mitosis, the identity of its mitotic substrates, and how it promotes cytokinesis. Overall this work will reveal how LATS and YAP are regulated by specific stimuli. These studies will have an important impact on our understanding of tumor suppression and tissue regeneration and may lead to better ways to manipulate these important processes.