Investigating of the Function of Copine A in Dictyostelium

盘基网柄菌中Copine A的功能研究

• 批准号: 7515414
• 负责人: CYNTHIA K DAMER
• 金额: $ 20.5万
• 依托单位: CENTRAL MICHIGAN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7515414
• 关键词: Affinity Chromatography; Amoeba genus; Animal Model; Apoptosis; Applications Grants; Binding; Binding Proteins; Biochemical; Blood; Bone Marrow Cells; C-terminal; C2 Domain; Calcium; Cell Differentiation process; Cell physiology; Cells; Chemotaxis; Condition; Development; Developmental Process; Dictyostelium; Dictyostelium discoideum; Disease; Eukaryotic Cell; Fruit; Genes; Goals; Human; Human Genome; Immune; In Vitro; Integrins; Knowledge; Lead; Leukocytes; Life; Localized; Location; Mediating; Membrane; Microscopic; Modeling; Molecular; Molecular Chaperones; Morphogenesis; N-terminal; Organism; Pattern; Phospholipids; Play; Protein Binding; Protein Binding Domain; Protein Family; Proteins; Public Health; Recruitment Activity; Reproduction spores; Role; Signal Pathway; Signal Transduction; Staging; Starvation; Structure; Techniques; Testing; cell motility; cell type; copine; design; extracellular; in vivo; mutant; novel; programs; protein function; research study; response; yeast two hybrid system

DESCRIPTION (provided by applicant): Copines are highly conserved calcium-dependent membrane-binding proteins found in numerous diverse eukaryotic organisms, including humans. Studies with human copines suggest that copines may act in signaling pathways by binding to target proteins and then recruiting these target proteins to a particular membrane in response to an increase in calcium concentration. This proposal describes experiments to specifically test the above hypothesis using developing Dictyostelium as a model. Dictyostelium can live independently as single celled amoebae, but when placed in starvation conditions, single amoeba signal each other to first aggregate and then differentiate into cells that form a multicellular fruiting body. This simple developmental program in Dictyostelium is widely used to study not only development, but also several basic cell processes including chemotaxis-mediated cell motility, signal transduction, programmed cell death cell, and cell differentiation. Previous studies show that one of the copine proteins in Dictyostelium, CpnA, is required for normal development and the developmental expression patterns of several of the other copine genes suggest they too may be regulators of development. The experiments outlined in specific aim 1 are designed to further characterize the role of CpnA in development as well as investigate the function of two other copine proteins. These experiments will allow us to determine the developmental and cellular processes in which copines function; however, the exact mechanism that they use to carry these functions will require biochemical studies like the ones proposed in the second specific aim. Experiments outlined in specific aim 2 are designed to identify the target proteins of the Dictyostelium copines to explore the mechanistic role of copines as chaperone or recruiting proteins functioning in signaling pathways. The human genome has eight copine genes and most of them are expressed in bone marrow cells and blood leukocytes; therefore, copines are potentially involved in cell signaling pathways within these cells. Increasing our scientific knowledge of how copines function could lead to a better understanding of human blood and immune cell-related diseases. PUBLIC HEALTH RELEVANCE: The proposal outlines experiments to investigate the functions of a novel family of proteins called copines. Copines are hypothesized to have roles in cell signaling pathways, but their exact functions are unknown. The human genome has eight copine genes and most are found in bone marrow cells and blood leukocytes. Increasing our scientific knowledge of how copines function could potentially lead to a better understanding of human blood and immune cell related diseases.

描述（由申请人提供）：Copines是高度保守的钙依赖性膜结合蛋白，存在于许多不同的真核生物中，包括人类。对人类copines的研究表明，copines可能通过与靶蛋白结合，然后将这些靶蛋白募集到特定的膜上，以响应钙浓度的增加，从而在信号通路中起作用。本提案描述了以发展盘基骨为模型具体检验上述假设的实验。盘基骨柱可以作为单细胞变形虫独立生活，但当处于饥饿状态时，单个变形虫相互发出信号，首先聚集，然后分化成形成多细胞子实体的细胞。Dictyostelium的这个简单的发育程序不仅被广泛用于研究发育，而且被广泛用于研究一些基本的细胞过程，包括趋化介导的细胞运动、信号转导、细胞程序性死亡细胞和细胞分化。先前的研究表明盘基骨菌中的一种copine蛋白CpnA是正常发育所必需的，其他几个copine基因的发育表达模式表明它们也可能是发育的调节因子。在特定目标1中概述的实验旨在进一步表征CpnA在发育中的作用，并研究另外两种copine蛋白的功能。这些实验将使我们能够确定复制体发挥作用的发育和细胞过程；然而，它们用来实现这些功能的确切机制将需要生化研究，就像第二个具体目标中提出的那样。在特定目标2中概述的实验旨在鉴定copines盘基骨菌的靶蛋白，以探索copines作为伴侣或招募蛋白在信号通路中的机制作用。人类基因组有8个copine基因，其中大部分在骨髓细胞和血液白细胞中表达；因此，复制因子可能参与这些细胞内的细胞信号通路。增加我们对复制细胞如何发挥作用的科学知识，可以更好地了解人类血液和免疫细胞相关疾病。公共卫生相关性：该提案概述了研究一种称为复制蛋白的新蛋白质家族功能的实验。复制基因被假设在细胞信号通路中起作用，但其确切功能尚不清楚。人类基因组有8个copine基因，大多数存在于骨髓细胞和血液白细胞中。增加我们对复制细胞功能的科学知识可能会更好地理解人类血液和免疫细胞相关疾病。