CpnA's role in cAMP signaling and the actin cytoskeleton in Dictyostelium

CpnA 在盘基网柄菌 cAMP 信号传导和肌动蛋白细胞骨架中的作用

• 批准号: 8434437
• 负责人: CYNTHIA K DAMER
• 金额: $ 31.4万
• 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 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8434437
• 关键词: Actins; Affect; Affinity Chromatography; Amoeba genus; Animal Model; Apoptosis; Applications Grants; Award; Binding; Binding Proteins; Biochemical; Biochemical Genetics; Calcium; Cell Culture Techniques; Cell Differentiation process; Cell Line; Cell physiology; Cells; Chemotaxis; Cyclic AMP; Cytoskeleton; Data; Defect; Development; Developmental Process; Dictyostelium; Dictyostelium discoideum; Disease; Eukaryotic Cell; Fruit; Genes; Genome; Goals; Grant; Heating; Human; Human Genome; Immune; Immune response; Immunoprecipitation; In Vitro; Knowledge; Lead; Life; Light; Malignant Neoplasms; Mediating; Membrane; Microfilaments; Microscopy; Molecular; Morphogenesis; Organism; Phenotype; Play; Protein Binding; Protein Family; Proteins; Proteolysis; Recruitment Activity; Regulation; Reproduction spores; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Starvation; Structure; System; Techniques; Testing; Time; Tumor Necrosis Factor Receptor; Yeasts; cell motility; cell type; copine; depolymerization; design; extracellular; genetic regulatory protein; in vivo; mutant; novel; phosphoric diester hydrolase; polymerization; programs; protein function; public health relevance; receptor; research study; slug; transcription factor; yeast two hybrid system

DESCRIPTION (provided by applicant): Copines are highly conserved calcium-dependent membrane-binding proteins found in numerous diverse eukaryotic organisms. The wide array of organisms ranging from single-celled organisms to humans in which copines are found suggest that copines carry out fundamental functions important in most eukaryotic cells. A growing body of evidence indicates that copines act as regulatory proteins in signaling pathways. The model organism Dictyostelium discoideum has six copine genes (cpnA-cpnF) and provides an ideal system for studying copine function. 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, and cell differentiation. Previous studies showed that one of the copine proteins in Dictyostelium, CpnA, is required for normal development. The overall goal of the previous grant award was to correlate our findings on cpnA- cells during developmental processes (aim 1) and the identification of the target proteins of CpnA (aim 2) to generate more specific hypotheses about the function of CpnA. Significant progress was made on both of these specific aims and data collected during the previous grant award period has led to two new hypotheses about the function of CpnA. These two hypotheses form the two specific aims of this renewal proposal: 1) Determine if CpnA is a negative regulator of the cAMP phosphodiesterase, RegA and 2) Determine if CpnA is a negative regulator of actin filament polymerization. This grant proposal describes experiments using biochemical, genetics, and microscopy approaches to test these hypotheses. The results of these experiments will make a significant impact on this field by being the first to define a mechanistic role for a copine protein. Human copines have been implicated to play a role in cancer by regulating signaling pathways involving the ErB2 receptor and in immune responses by regulating signaling pathways involving the TNF (tumor necrosis factor) receptor. Increasing our scientific knowledge of how copines function could lead to a better understanding of cancer and immune cell-related diseases.

描述（由申请人提供）：COPINE是在许多多种真核生物中发现的高度保守钙依赖性的膜结合蛋白。从单细胞生物到人类的各种生物都发现，发现章节表明，在大多数真核细胞中，cosine具有重要的基本功能。越来越多的证据表明，在信号通路中，封索是调节蛋白的作用。模型生物体distelium discoideum具有六个Copine基因（CPNA-CPNF），并提供了研究美联体功能的理想系统。 Dictyostelium可以独立地作为单细胞变形虫生活，但是当放置在饥饿条件下时，单个变形虫相互信号以首先聚集，然后分化成形成多细胞水果体的细胞。这个简单的开发程序在Dictyostelium中不仅用于研究发育，还用于研究几种基本细胞过程，包括趋化性介导的细胞运动，信号转导，程序性细胞死亡和细胞分化。先前的研究表明，正常发育中需要一种cpna中的一种cpna的蛋白质。先前赠款奖的总体目标是将我们在发育过程中CPNA细胞的发现（AIM 1）和CPNA目标蛋白（AIM 2）的鉴定相关联（AIM 2），以产生有关CPNA功能的更具体的假设。在上一个赠款期间收集的这些特定目标和收集的数据都取得了重大进展，这导致了两个关于CPNA功能的新假设。这两个假设构成了该更新建议的两个具体目的：1）确定CPNA是否是CAMP磷酸二酯酶的负调节剂，REGA和2）确定CPNA是否是肌动蛋白丝聚合的负调节剂。该赠款提案描述了使用生化，遗传学和显微镜方法检验这些假设的实验。这些实验的结果将是第一个定义美联体蛋白机械作用的人，从而对该领域产生重大影响。通过调节涉及ERB2受体的信号传导途径和免疫反应，通过调节涉及TNF（肿瘤坏死因子）受体的信号通路来调节涉及ERB2受体的信号通路和免疫反应，这与人类的合同有关。提高我们对Copines功能的科学知识可能会导致对癌症和免疫细胞相关疾病的更好理解。