A Novel Mechanism of GTPase Regulation: Arl2 Interaction with TBCD

GTPase 调节的新机制：Arl2 与 TBCD 的相互作用

• 批准号: 8783663
• 负责人: Joshua W Francis
• 金额: $ 4.27万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8783663
• 关键词: ADP-Ribosylation Factors; Binding; Biochemical; Biological; Biological Assay; Cardiac; Cell Proliferation; Cell division; Cell physiology; Cells; Centrosome; Complex; Cytosol; Data; Development; Disease; Dissociation; Dominant-Negative Mutation; Embryo; Energy Metabolism; Eukaryotic Cell; Family; Goals; Guanine Nucleotide Exchange Factors; Guanosine Triphosphate Phosphohydrolases; Health; Heart Diseases; Human; Hypoxia; In Vitro; Lead; Learning; Life; Link; Maintenance; Malignant Neoplasms; Mammalian Cell; Mammary Gland Parenchyma; Microtubule Polymerization; Microtubules; Mitotic spindle; Modeling; Molecular Chaperones; Nucleotides; Pancreas; Pathway interactions; Phenotype; Play; Polymers; Positioning Attribute; Preparation; Process; Property; Protein Binding; Proteins; Recombinants; Regulation; Retinal Degeneration; Role; Series; Signal Transduction; Solid Neoplasm; Source; System; Testing; Transcription Coactivator; Tubulin; Vascularization; Warburg Effect; base; cell growth; cofactor; design; human disease; insight; kidney cell; member; milligram; mutant; novel; overexpression; public health relevance; research study; tumor growth

DESCRIPTION (provided by applicant): The strict regulation of energy metabolism and cell proliferation is critical for the normal function of all eukaryotic life. Improper regulation of thse fundamental cellular processes is associated with multiple disease states, including heart disease and cancer. Examples include shifting the sources of ATP in cells as a result of the Warburg effect or hypoxia, resulting from solid tumor growth exceeding vascularization or cardiac insufficiency. ADP-ribosylation factor like-2 (Arl2), a member of the Arf family of regulatory GTPases, is an ancient and ubiquitous eukaryotic cell regulator that is involved in the maintenance of cellular ATP levels, the progression of cell division and the regulation of transcriptional activators involved in cell proliferation. Maintaining the necessary levels of activated Arl2 within a cell is crucial for the proper regulation of these essential processes. However, the biological mechanisms controlling Arl2 activity are unknown. The majority of cellular Arl2 is in a cytosolic complex with the tubulin-specific co-chaperone, cofactor D (TBCD). We will perform in vitro biochemical analyses to test the hypothesis that dissociation from TBCD is a critical, regulated step in Arl2 function in cells. Additionally, we will test the model that Arl2/TBCD is also an active complex required for proper regulation of centrosomal functions and microtubule dynamics. In specific aim 1, we will utilize recombinant preparations of Arl2 and TBCD to identify the mechanisms regulating assembly/disassembly of the Arl2/TBCD complex. In specific aim 2, we will use cell-based assays to determine the importance of Arl2 and TBCD interaction in regulating centrosomal and microtubule functions. The results of this study will provide fundamental groundwork in our long-term goal of explaining the mechanistic roles of Arl2 as a ubiquitous regulator that is indispensable to the regulation of several essential cellula processes. Further insight into the mechanisms underlying Arl2 regulation will ultimately lead to a better understanding of fundamental eukaryotic functions, such as energy metabolism and cell proliferation.

描述（申请人提供）：严格调节能量代谢和细胞增殖对所有真核生物的正常功能至关重要。这些基本细胞过程的不当调节与多种疾病状态有关，包括心脏病和癌症。例如，实体瘤生长超过血管化或心功能不全导致的Warburg效应或缺氧导致细胞中ATP来源的改变。adp -核糖基化因子样-2 （Arl2）是调节gtpase的Arf家族成员，是一种古老而普遍存在的真核细胞调节剂，参与细胞ATP水平的维持、细胞分裂的进展以及参与细胞增殖的转录激活因子的调节。在细胞内维持必要水平的活化Arl2对于这些基本过程的适当调节至关重要。然而，控制Arl2活性的生物学机制尚不清楚。大多数细胞Arl2与微管蛋白特异性的辅因子D （TBCD）在胞质复合体中。我们将进行体外生化分析，以验证从TBCD分离是Arl2在细胞中功能的关键调控步骤的假设。此外，我们将测试Arl2/TBCD也是中心体功能和微管动力学适当调节所需的活性复合物的模型。在具体目标1中，我们将利用Arl2和TBCD的重组制备来确定Arl2/TBCD复合物组装/拆卸的调节机制。在特定目标2中，我们将使用基于细胞的测定来确定Arl2和TBCD相互作用在调节中心体和微管功能中的重要性。这项研究的结果将为我们的长期目标提供基础基础，即解释Arl2作为一种无处不在的调节剂的机制作用，这种调节剂对几个基本细胞过程的调节是必不可少的。对Arl2调控机制的进一步了解将最终有助于更好地理解真核生物的基本功能，如能量代谢和细胞增殖。