P68 and Ca-calmodulin interaction in cell migration

P68 和 Ca-钙调蛋白在细胞迁移中的相互作用

• 批准号: 8693188
• 负责人: Zhi-Ren Liu
• 金额: $ 23.31万
• 依托单位: GEORGIA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8693188
• 关键词: ATP phosphohydrolase; Abnormal Cell; Accounting; Address; Affect; Animal Cancer Model; Binding; Binding Sites; Biological Assay; Biological Process; Calcium/calmodulin-dependent protein kinase; Calmodulin; Cell Nucleus; Cell physiology; Cells; Characteristics; Chimeric Proteins; Clinical; Complex; Cytoplasm; Cytoskeleton; Development; Disease; Embryonic Development; Epithelial Cells; Family; Goals; Guanosine Triphosphate Phosphohydrolases; Immune response; In Vitro; Injection of therapeutic agent; Knowledge; Lead; Length; Lymphocyte; Maps; Membrane; Microtubules; Molecular; Molecular Conformation; Motor; Motor Activity; Mus; Mutation; Neoplasm Metastasis; New Agents; Peptide Conformation; Peptides; Phosphorylation; Phosphorylation Site; Phosphotransferases; Play; Process; Protein Dephosphorylation; Protein Kinase; Protein Phosphatase 2A Regulatory Subunit PR53; Proteins; RNA; Research Project Grants; Role; Signal Transduction; Site; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Stimulus; Structural Models; Structure; System; Testing; Tissues; Wound Healing; base; cell motility; crosslink; disease diagnosis; migration; outcome forecast; peptide A; protein p68; public health relevance; research study; rho; tumor

DESCRIPTION (provided by applicant): Cell migration is an essential characteristic of cells in numerous fundamental biological processes, such as embryogenesis, tissue development, wound healing, and migration of lymphocytes in the immune response system. The molecular mechanism that governs this important cellular process is extensively studied. However, despite intensive characterizations, several important knowledge gaps remain, especially regarding the functional role of the key player, Ca-calmodulin (CaM), in the cell migration process. We have observed that the p68-CaM interaction is necessary for epithelial cell migration. Upon cell migration stimulation, the p68-CaM interaction was substantially enhanced. Binding to CaM caused localization of p68 from the cell nucleus to the cytoplasm. In the migrating cells, p68 and CaM co-localized to the migration leading edges (lamellipodia or fillopodia). Mutations that abolished p68 ATPase activity blocked the localization of CaM to the leading edge of migrating cells. Expression of a p68-calmodulin fusion protein led to the formation of lamellipodium and cell morphological changes and dramatically increased cell motility. The immunopurified p68-CaM fusion protein interacted with the cytoskeleton microtubule in vitro. In addition, a peptide derived from p68 IQ motifs strongly inhibited cell migration. As a consequence, treatment of tumor bearing mouse with the IQ peptide almost completely abolished cancer metastasis. Our observations suggest that the p68-CaM interaction is a new player in regulating the cell migration. The goal of this proposed research project is to understand the role of the p68-CaM interaction in cell migration process. We propose three specific aims to investigate the functional significance of p68-CaM interaction in cell migration. In specific aim 1, we will elucidate the mechanism that regulates the p68-CaM interaction. We will test our hypothesis that cell migration signals trigger phosphorylation of p68, which subsequently lead to the p68-CaM interaction. In specific aim 2, we will unravel the function of p68 at migration leading edge. In specific aim 3, we plan to focus on addressing the molecular mechanism that governors the substrate selection of p68 in different cellular processes. We believe that our results will ultimately be applied to develop new strategies for diseases diagnosis/prognosis and treatments.

描述（由申请人提供）：细胞迁移是许多基本生物学过程中细胞的重要特征，例如胚胎发生，组织发育，伤口愈合和免疫反应系统中淋巴细胞的迁移。广泛研究了控制这一重要细胞过程的分子机制。然而，尽管进行了密集的特征，但仍然存在一些重要的知识差距，尤其是关于关键参与者CA-Calmodulin（CAM）在细胞迁移过程中的功能作用。我们已经观察到P68-CAM相互作用对于上皮细胞迁移是必需的。细胞迁移刺激后，P68-CAM相互作用大大增强。与CAM的结合导致p68从细胞核定位到细胞质。在迁移的细胞中，p68和cam共定位于迁移前缘（lamellipodia或fillopodia）。废除P68 ATPase活性的突变阻塞了CAM的定位到迁移细胞的前缘。 p68-钙调蛋白融合蛋白的表达导致形成层状脂肪和细胞形态变化，并大大增加了细胞运动。免疫疗法的p68-CAM融合蛋白在体外与细胞骨架微管相互作用。另外，衍生自P68 IQ基序的肽强烈抑制细胞迁移。因此，用智商肽对肿瘤轴承的治疗几乎完全废除了癌症转移。我们的观察结果表明，P68-CAM相互作用是调节细胞迁移的新玩家。该提出的研究项目的目的是了解p68-CAM相互作用在细胞迁移过程中的作用。我们提出了三个特定的目的来研究功能 p68-CAM相互作用在细胞迁移中的重要性。在特定的目标1中，我们将阐明调节p68-CAM相互作用的机制。我们将测试我们的假设，即细胞迁移信号触发p68的磷酸化，后来导致p68-CAM相互作用。在特定的目标2中，我们将在迁移前缘揭示p68的功能。在特定目标3中，我们计划专注于解决不同细胞过程中p68底物选择的分子机制。我们认为，我们的结果最终将用于制定疾病的新策略诊断/预后和治疗。