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: 9804133
• 负责人: RICHARD E CHENEY
• 金额: $ 48.14万
• 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/9804133
• 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; 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; outcome forecast; 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.

项目总结/摘要 丝状伪足是质膜的指状延伸，允许细胞与它们的细胞接触并相互作用。 在诸如神经生长、血管形成和癌细胞扩散的过程中，环境受到影响。 丝状伪足尖端的一个知之甚少的结构（丝状伪足尖端复合体）构成了 丝状伪足肌动蛋白聚合、粘附和信号传导。我们对肌球蛋白X（Myo 10）的研究表明，它是一种 它是丝状伪足尖端复合体的主要成分，最近发现它在哺乳动物中具有重要作用。 在神经管闭合、眼睛形成、血管发育和色素沉着中的发育。患者 小眼症（眼睛异常小）最近被证明缺乏Myo 10，强烈表明突变 Myo 10中的基因可能导致人类隐性遗传病。越来越多的论文也表明，Myo 10 是癌细胞侵袭、转移和分裂中的关键蛋白，并且在癌细胞的主要分化中经常上调。 人类癌症这些和其他观察结果使Myo 10成为抗癌研究的一个有吸引力的靶点， 因此，确定Myo 10和丝状伪足尖端复合体的基本细胞生物学功能是必要的。 虽然丝状伪足尖端可以形成专门的粘连部位，但它们缺乏局灶性粘连的核心成分。 事实上，所有的丝状伪足尖端都含有Myo 10，这是一种我们已经证明可以将肌动蛋白与β-整合素连接起来的蛋白质， 细胞粘附分子。因为尖端复杂的组装和组成以及它如何随着它的变化而变化， 从伸展转换为回缩或粘附不清楚，我们将： 1)确定在起始、伸展、回缩和牵拉过程中丝状伪足尖端复合体的组成， 粘附这一目标将定义丝状伪足尖端复合体的组装途径和状态， 核心丝状伪足尖端组分包括Myo 10、其假定的相互作用物VASP和加帽蛋白。 2)确定有丝分裂回缩纤维尖端的粘连成分。虽然撤回 纤维在有丝分裂过程中具有锚定细胞的生物学关键作用，但对纤维的生物学特性几乎一无所知。 其尖端的粘连物的组成。Myo 10是已知的少数几种定位于细胞尖端的蛋白质之一。 收缩纤维，所以我们将使用它来研究这个知之甚少，但细胞粘附的关键部位。 3)探讨基底外侧丝状伪足的组成及功能。我们发现，在 极化上皮细胞Myo 10靶向基底外侧表面上的丝状伪足尖端，而不是基底外侧表面上的丝状伪足尖端。 相同细胞上的顶端微绒毛。由于Myo 10提供了一个探针，用于这种在很大程度上未表征的尖端， 类的丝状伪足，我们将用它来研究这些结构的基本细胞生物学。这项研究 将定义由Myo 10和尖端复合物标记的三个知之甚少的结构的基本性质： 丝状伪足尖端粘连，回缩纤维粘连，以及两极基底外侧表面上的丝状伪足 上皮细胞