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基本特征，如胚胎发生、组织发育、伤口愈合和免疫应答系统中淋巴细胞的迁移。管理这一重要细胞过程的分子机制被广泛研究。然而，尽管密集的表征，一些重要的知识差距仍然存在，特别是关于关键球员，钙-钙调蛋白（CaM），在细胞迁移过程中的功能作用。我们已经观察到，p68-CaM相互作用是上皮细胞迁移所必需的。在细胞迁移刺激后，p68-CaM相互作用显著增强。结合到钙调素引起的本地化的p68从细胞核到细胞质。在迁移细胞中，p68和CaM共定位于迁移前缘（板状伪足或丝状伪足）。突变，废除p68 ATP酶活性的本地化钙调素的迁移细胞的前沿。p68-钙调蛋白融合蛋白的表达导致板状伪足的形成和细胞形态学的变化，并显着增加细胞运动。免疫纯化的p68-CaM融合蛋白在体外与细胞骨架微管相互作用。此外，来自p68 IQ基序的肽强烈抑制细胞迁移。因此，用IQ肽治疗荷瘤小鼠几乎完全消除了癌症转移。我们的观察表明，p68-CaM相互作用是一个新的球员在调节细胞迁移。本研究的目的是了解p68-CaM相互作用在细胞迁移过程中的作用。我们提出了三个具体的目标，以调查功能 p68-CaM相互作用在细胞迁移中意义在具体目标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磷酸化，随后导致p68-CaM相互作用。在具体目标2中，我们将阐明p68在迁移前沿的功能。在具体的目标3中，我们计划专注于解决在不同的细胞过程中调控p68底物选择的分子机制。我们相信，我们的研究结果最终将应用于开发疾病诊断/预后和治疗的新策略。