The filopodial tip complex in adhesion, migration, and signaling

丝状伪足尖端复合体在粘附、迁移和信号传导中的作用

• 批准号: 10216311
• 负责人: RICHARD E CHENEY
• 金额: $ 43.07万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-02 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10216311
• 关键词: Actins; Adhesions; Apical; Binding; Biological; Biological Process; Brain; Cancer Biology; Cell Adhesion; Cell membrane; Cells; Cellular biology; Chemotaxis; Child; Complex; Defect; Development; Disease; Embryonic Development; Epithelial Cells; Eye; Fiber; Filopodia; Fingers; Focal Adhesions; Genetic Diseases; Growth; Health; Heterogeneity; Human; Integrin beta Chains; Integrins; Knock-out; Link; Malignant Neoplasms; Malignant neoplasm of lung; Mammalian Cell; Microphthalmos; Mitosis; Mitotic; Molecular; Mus; Mutation; Myosin ATPase; Neoplasm Metastasis; Nerve; Neural Tube Closure; Paper; Pathway interactions; Patients; Physiological; Pigmentation physiologic function; Process; Prognosis; Property; Proteins; Reporting; Research; Resolution; Role; Signal Transduction; Site; Structure; Surface; Tail; Work; adhesion receptor; anticancer research; blood vessel development; cancer cell; cell motility; cellular microvillus; eye blood vessel; eye formation; human disease; live cell imaging; malignant breast neoplasm; melanoma; migration; monolayer; paxillin; polymerization; recessive genetic trait; structural biology; vasodilator-stimulated phosphoprotein; virtual

Project Summary/Abstract Filopodia are finger-like extensions of the plasma membrane that allow cells to contact and interact with their surroundings in processes such as nerve growth, blood vessel formation, and the spread of cancer cells. A poorly understood structure at the tips of filopodia, the filopodial tip complex, constitutes a key site of filopodial actin polymerization, adhesion, and signaling. Our research with myosin-X (Myo10) shows that it is a major component of the filopodial tip complex and recently revealed that it has important roles in mammalian development in neural tube closure, eye formation, blood vessel development, and pigmentation. A patient with microphthalmia (unusually small eyes) was recently shown to lack Myo10, strongly suggesting that mutations in Myo10 can cause recessive genetic disease in humans. A growing number of papers also show that Myo10 is a key protein in the invasion, metastasis, and division of cancer cells and is frequently upregulated in major human cancers. These and other observations make Myo10 an attractive target for anticancer research and make it essential to determine the fundamental cell biological functions of Myo10 and filopodial tip complex. Although filopodial tips can form specialized sites of adhesion, they lack core components of focal adhesions. Virtually all filopodial tips contain Myo10, a protein that we have shown can link actin to β-integrins, key molecules in cell adhesion. Because the assembly and composition of the tip complex and how it changes as it converts from extension to retraction or adhesion is not understood, we will: 1) Determine the composition of the filopodial tip complex during initiation, extension, retraction, and adhesion This aim will define the assembly pathway and states of the filopodia tip complex with respect to core filopodial tip components including Myo10, its putative interactor VASP, and capping protein. 2) Define the composition of the adhesions at the tips of mitotic retraction fibers. Although retraction fibers have the biologically crucial role of anchoring cells during mitosis, almost nothing is known about the composition of the adhesions at their tips. Myo10 is one of the few proteins known to localize to the tips of retraction fibers, so we will use it to investigate this poorly understood, but key site of cell adhesion. 3) Investigate the composition and functions of basolateral filopodia. We have discovered that in polarized epithelial cells Myo10 is targeted to the tips of filopodia on the basolateral surface rather than the apical microvilli on the same cells. Because Myo10 provides a probe for the tips of this largely uncharacterized class of filopodia, we will use it to investigate the basic cell biology of these structures. Together this research will define the basic properties of three poorly understood structure marked by Myo10 and the tip complex: filopodial tip adhesions, retraction fiber adhesions, and the filopodia on the basolateral surfaces of polarized epithelial cells.

项目概要/摘要 丝状伪足是质膜的手指状延伸，允许细胞与其接触并相互作用 神经生长、血管形成和癌细胞扩散等过程中的周围环境。 丝状伪足尖端的一个人们知之甚少的结构，即丝状伪足尖端复合体，构成了丝状伪足的一个关键位点。 丝状肌动蛋白聚合、粘附和信号传导。我们对肌球蛋白-X (Myo10) 的研究表明，它是 丝状足尖端复合体的主要组成部分，最近发现它在哺乳动物中具有重要作用 神经管闭合、眼睛形成、血管发育和色素沉着的发育。一名患者患有 最近发现小眼症（异常小的眼睛）缺乏 Myo10，强烈表明突变 Myo10中的Myo10可导致人类隐性遗传病。越来越多的论文也表明 Myo10 是癌细胞侵袭、转移和分裂的关键蛋白，在主要细胞中经常上调 人类癌症。这些和其他观察结果使 Myo10 成为抗癌研究的有吸引力的目标 因此确定 Myo10 和丝状尖端复合体的基本细胞生物学功能至关重要。 尽管丝状伪足尖端可以形成专门的粘附位点，但它们缺乏粘着斑的核心成分。 几乎所有的丝状足尖端都含有 Myo10，我们已经证明这种蛋白质可以将肌动蛋白与 β-整合素连接起来，这是关键 细胞粘附分子。因为尖端的组装和组成非常复杂，并且随着它的变化而变化 从延伸转换为收缩或粘附不明白，我们将： 1) 确定丝状伪足尖端复合体在起始、伸展、收缩和收缩过程中的组成 粘附 该目标将定义丝状伪足尖端复合体的组装途径和状态 核心丝状伪足尖端成分包括 Myo10、其假定的相互作用蛋白 VASP 和加帽蛋白。 2) 定义有丝分裂回缩纤维尖端粘连的成分。虽然撤回 纤维在有丝分裂过程中具有锚定细胞的重要生物学作用，但人们对纤维几乎一无所知 尖端粘连的成分。 Myo10 是已知定位于末端的少数蛋白质之一 回缩纤维，因此我们将用它来研究这个知之甚少但关键的细胞粘附位点。 3）研究基底外侧丝状伪足的组成和功能。我们发现，在 极化上皮细胞 Myo10 的目标是基底外侧表面的丝状伪足的尖端，而不是基底外侧的丝状伪足的尖端。 相同细胞上的顶端微绒毛。因为 Myo10 为这种基本上没有特征的尖端提供了探针 丝状伪足类，我们将用它来研究这些结构的基本细胞生物学。共同开展这项研究 将定义由 Myo10 和尖端复合物标记的三个鲜为人知的结构的基本属性： 丝状伪足尖端粘连、回缩纤维粘连以及偏振基底外侧表面的丝状伪足 上皮细胞。