Collaboration between actin nucleators - Spire and Cappuccino

肌动蛋白成核剂 - Spire 和 Cappuccino 之间的合作

• 批准号: 9185648
• 负责人: Margot E Quinlan
• 金额: $ 30.34万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9185648
• 关键词: Actins; Animal Genetics; Animals; Binding; Biological Assay; Boxing; Cell Polarity; Cell division; Cells; Collaborations; Cytoplasm; Cytoskeleton; Data; Development; Drops; Drosophila genus; Drosophila inturned protein; Epithelial Cells; Excision; Fertility; Filament; Fluorescence; Fluorescence Microscopy; Fluorescence Resonance Energy Transfer; Genetic; Geometry; Goals; Health; Homologous Gene; Human; In Vitro; Knowledge; Lead; Learning; Life; Mammals; Measurement; Membrane; Microfilaments; Microtubules; Modeling; Molecular; Mus; Mutagenesis; Neurons; Oocytes; Oogenesis; Pathway interactions; Positioning Attribute; Process; Proteins; Proteomics; Role; Staging; Stream; Structure; Testing; Time; Work; base; cell motility; egg; in vivo; operation; protein degradation; reconstitution; reverse genetics; single molecule

Project Summary The cytoskeleton is essential to many aspects of life, cell division and motility being prime examples. The overarching goal of this proposal is to understand the roles of two actin nucleators, Spire (Spir) and Cappuccino (Capu). We are studying these protein in Drosophila, where they are known to be essential to polarity establishment during early development. Spir and Capu build an actin mesh that traverses the Drosophila oocyte throughout mid-oogenesis. The presence of the mesh and its removal at the correct developmental stage are both critical to polarity establishment. Thus learning how Spir and Capu collaborate to build the mesh is essential to a fundamental understanding of how they contribute to polarity establishment during development. In the proposed work, we will focus on two aspects of Spir-Capu operation: 1) the mechanism by which they assemble actin filaments and 2) how Spir and Capu are spatially and temporally regulated, in order to establish the actin mesh and then remove it when appropriate. We have developed a working model in which Spir and Capu are both active throughout mid-oogenesis (stages 5-9); Spir enhances nucleation by Capu, specifically at membranes; together they build a dynamic actin mesh until stage 10 when Spir and Capu protein levels drop, the mesh disappears, and streaming begins. We will test this model using both in vitro and in vivo approaches with assays that span scales from single molecule fluorescence microscopy to whole animal genetics. Testing this model will lead to a mechanistic understanding of two proteins, conserved throughout metazoan species. It will advance our knowledge of the cytoskeleton and how it is controlled. Given the co-existence of this pair of proteins in polar cells, including neurons and epithelial cells in mammals, it has been proposed that their role as polarity factors in Drosophila is conserved. Thus we anticipate that what is learned about Spir and Capu in Drosophila oogenesis will be applicable to our understanding of the cytoskeleton, cell polarity, fertility, development and health in many animals, ranging from Drosophila to humans.

项目摘要 细胞骨架对生命的许多方面都是必不可少的，细胞分裂和运动就是最好的例子。这个 这项建议的首要目标是了解两个肌动蛋白核仁的作用，Spire(Spir)和 卡布奇诺(Capu)。我们正在研究果蝇体内的这些蛋白质，已知它们是果蝇 在早期发育过程中的极性建立。Spir和Capu构建了一个肌动蛋白网格，该网格遍历 果蝇卵母细胞贯穿卵子发生中期。网格的存在及其在正确位置的移除 发育阶段对两极的建立都是至关重要的。从而了解Spir和Capu如何合作 建立网络对于从根本上理解它们对极性建立的贡献是必不可少的 在开发过程中。在拟议的工作中，我们将重点关注Spir-Capu运营的两个方面：1) 它们组装肌动蛋白细丝的机制以及2)Spir和Capu在空间和时间上是如何 调节，以便建立肌动蛋白网状结构，然后在适当的时候移除它。我们已经开发出一种 在整个卵子发生中期(阶段5-9)，Spir和Capu都活跃的工作模式；Spir增强 由Capu成核，特别是在膜上；它们共同构建动态肌动蛋白网状结构，直到阶段10 Spir和Capu蛋白水平下降，网状结构消失，流开始。我们将使用以下工具测试此模型 无论是在体外还是在体内，分析方法都跨越了单分子荧光的范围 从显微镜到整个动物遗传学。测试这个模型将导致对两个模型的机械性理解 蛋白质，保存在后生动物物种中。它将促进我们对细胞骨架的了解以及如何 它是被控制的。鉴于这对蛋白质在极地细胞中的共存，包括神经元和上皮细胞 在哺乳动物中，细胞作为极性因子在果蝇中的作用是保守的。因此，我们 预计有关Spir和Capu在果蝇卵子发生中的知识将适用于我们的 对许多动物的细胞骨架、细胞极性、生育、发育和健康的了解，从 果蝇对人类的影响。