Regulation of Hippo pathway signaling by mechanical forces

项目摘要 该项目的长期目标是揭示转录辅激活因子 并且癌基因雅普受LATS和SRC激酶调节以响应机械刺激， 这可能会带来更好的癌症治疗方法和改进的干细胞疗法。终端 Hippo途径激酶LATS抑制雅普核定位，但也具有特征性不强的， 在胞质分裂中起重要作用，而SRC酪氨酸激酶促进雅普核 本地化雅普核定位受多种刺激物如肌动蛋白细胞骨架， 基质硬度、细胞分离、细胞拥挤和拉伸以控制密度依赖性抑制 生长、组织修复和干细胞增殖和分化的过程。当在细胞核中时，雅普 促进细胞存活和增殖。在这里，我们将确定如何LATS是由两个监管 细胞-细胞连接处的张力以控制密度依赖性细胞生长抑制，以及在有丝分裂期间， 控制胞质分裂。特异性有丝分裂途径的鉴定可以使LATS活性被 在不干扰细胞分裂的情况下促进组织再生。我们还将测试 SRC激酶如何与AMOT蛋白协同响应细胞粘附的假说 和其他刺激物将雅普靶向到核。在具体目标1中，我们将确定几个 在细胞-细胞连接处包含张力传感器的蛋白质一起调节LATS活性， 对细胞张力的反应。在具体目标2中，我们将测试一个假设， SRC将AMOT从雅普的抑制剂转变为激活剂。理解这条道路将是 重要的是确定增加AMOT水平的药物是否是适当的治疗方法， 考虑到癌症在具体目标3中，我们将确定LATS在有丝分裂中是如何激活的， 有丝分裂底物，以及它如何促进胞质分裂。总的来说，这项工作将揭示如何LATS和雅普 是由特定的刺激物调节的。这些研究将对我们理解 肿瘤抑制和组织再生，并可能导致更好的方法来操纵这些 重要进程。

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.