Microfilaments in Budding Yeast and the Nematode

出芽酵母和线虫中的微丝

• 批准号: 7609087
• 负责人: Anthony P. Bretscher
• 金额: $ 48.07万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-02-01 至 2011-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7609087
• 关键词: Actins; Address; Animals; Binding; Biochemical; Biological; Biological Assay; Biology; Caenorhabditis elegans; Cell Cycle; Cell Polarity; Cell division; Cells; Cellular biology; Complement; Cytoskeleton; Defect; Disease; Eukaryota; F-Actin; Family; Filament; Genetic; Genetic Screening; Grant; Growth; Investigation; Location; Microfilaments; Microtubules; Molecular; Morphogenesis; Mothers; Mutation; Myosin ATPase; Myosin Type V; Nematoda; Nuclear; Organelles; Organism; Pathway interactions; Pattern; Phosphatidylinositols; Phosphorylation; Play; Process; Property; Proteins; Regulation; Role; Route; Running; Saccharomycetales; Secretory Vesicles; Site; Structure; Syndrome; Time; Vacuole; Vertebrates; Yeasts; cell growth; cell motility; clinically relevant; deafness; insight; member; peroxisome; profilin; protein function; segregation; tool

DESCRIPTION (provided by applicant): Cells have evolved mechanisms to establish spatial order of their constituent organelles and provide direction for transport of cellular components. This is most apparent in the need for cell polarity and the ability to segregate organelles appropriately during cell division. We have been using the budding yeast to study how the actin cytoskeleton brings about this cellular organization. We have shown that bundles of polarized actin filaments, nucleated by formins and running parallel to the axis of cell division, provide the framework for polarized growth and organelle segregation. We and others have also shown that an essential myosin-V, whose heavy chain is encoded by MYO2, uses these cables to transport secretory vesicles for polarized growth, as well as many organelles, including microtubules for nuclear orientation, the TGN, the vacuole, peroxisomes, etc. for segregation during the cell cycle. In this renewal, we propose to extend our studies on yeast formins, as well as undertake a new project investigating the breadth of functions that formins perform in a simple multicellular organism, the nematode Caenorhabditis elegans. We shall also continue with our studies on Myo2p, the best-understood member of the highly conserved myosin-V family, to investigate how it selects, delivers and releases it cargos. Specifically, we propose to address the following questions: (1) How is the nucleating activity of yeast formins controlled in time and space? We propose to use genetic and biochemical approaches to identify factors that cooperate with the yeast formin Bnilp, and understand in biochemical terms how these function; (2) What is the range of structures that formins perform? The nematode has single representative formins for almost all the classes of formins common to metazoans. We shall initiate a project to study the location, biochemical properties and functions of these different formins; (3) What proteins function with Myo2p, and how does it select between cargos? Genetic and cell biological studies will be expanded to identify factors (including specific phosphoinositides) that function with Myo2p in cargo selection, in its regulation, and in cargo delivery. These studies will contribute to a fundamental understanding of how cellular order is generated. Defects in related proteins in vertebrates are associated with diseases, including mutations in formin DNFA1, resulting in non-syndromic deafness, as well as in myosin-Vs, for example in Griscelli's syndrome. Thus, the proposed studies will provide insight into a basic cell biology question as well as have clinical relevance.

描述（由申请人提供）：细胞已经进化出建立其组成细胞器的空间秩序的机制，并为细胞组分的运输提供方向。这是最明显的，在细胞分裂过程中需要细胞极性和适当分离细胞器的能力。我们一直在使用芽殖酵母来研究肌动蛋白细胞骨架如何产生这种细胞组织。我们已经表明，成束的极化肌动蛋白丝，成核的formins和运行平行于轴的细胞分裂，提供了框架的极化生长和细胞器分离。我们和其他人还表明，一个重要的肌球蛋白-V，其重链由MYO 2编码，使用这些电缆运输分泌囊泡的极化生长，以及许多细胞器，包括微管的核方向，TGN，空泡，过氧化物酶体等分离在细胞周期中。在这次更新中，我们建议扩展我们对酵母formins的研究，并开展一个新的项目，调查formins在一个简单的多细胞生物体，线虫秀丽隐杆线虫中执行的功能的广度。我们还将继续对Myo 2 p的研究，Myo 2 p是高度保守的肌球蛋白V家族中最好的成员，以研究它如何选择，递送和释放它的货物。具体而言，我们提出解决以下问题：（1）如何在时间和空间控制酵母formins的成核活性？我们建议使用遗传和生物化学的方法来确定因素，合作与酵母菌Bnilp，并了解在生化方面如何这些功能;（2）什么是范围内的结构，formins执行？该线虫具有单一的代表性形式，几乎代表了后生动物常见的所有形式。我们将启动一个项目，研究这些不同的formins的位置，生化特性和功能;（3）什么蛋白质与Myo 2 p的功能，它如何选择货物之间？遗传和细胞生物学研究将扩大到确定的因素（包括特定的磷酸肌醇）与Myo 2 p在货物选择，在其调节，并在货物交付功能。这些研究将有助于从根本上了解细胞秩序是如何产生的。脊椎动物中相关蛋白质的缺陷与疾病相关，包括导致非综合征性耳聋的DNFA 1突变，以及肌球蛋白-Vs突变，例如Griscelli综合征。因此，拟议的研究将提供对基本细胞生物学问题的深入了解，并具有临床意义。