Role of GTP-binding/transglutaminase in cell survival

GTP 结合/转谷氨酰胺酶在细胞存活中的作用

基本信息

批准号：
7220564
负责人：
RICHARD A. CERIONE
金额：
$ 25.47万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-06-01 至 2008-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7220564
关键词：
1-Phosphatidylinositol 3-Kinase 70-kDa Ribosomal Protein S6 Kinases Amyloid Apoptosis Apoptotic Architecture Binding Binding Sites Biochemical Genetics Biogenic Amines Biological Bone Development Cardiac Cell Survival Cell physiology Cells Cellular Stress Complex Condition Coupled Data Development Dominant-Negative Mutation Ensure Equilibrium Event Exhibits Funding G-substrate GTP Binding GTP-Binding Proteins Glutamine Goals Guanine Nucleotides Guanosine Diphosphate Guanosine Triphosphate HL-60 Cells Hydrolysis Lead Light Link Lysine Mediating Molecular Mus Mutation NGFR Protein Natural regeneration Nerve Growth Factor Receptors Nerve Growth Factors Neurons Nucleotides Numbers Outcome PC12 Cells Pathway interactions Pheochromocytoma Play Polyamines Protein Binding Proteins Range Rattus Reaction Regulation Research Personnel Retinoblastoma Genes Retinoblastoma Protein Roentgen Rays Role Running Series Signal Pathway Signal Transduction Signaling Protein Stimulus Structure System Testing Tissues Transcriptional Activation Transferase Transglutaminases Tretinoin Up-Regulation Work aluminum fluoride amino group axon growth base cdc42 GTP-Binding Protein cell growth covalent bond cytotoxic design eIF-5A extracellular glutamine transport protein insight interest mouse Gdi2 protein mutant polymerization prevent programs protein function receptor coupling response structural biology three dimensional structure

1-Phosphatidylinositol 3-Kinase 70-kDa Ribosomal Protein S6 Kinases Amyloid Apoptosis Apoptotic Architecture Binding Binding Sites Biochemical Genetics Biogenic Amines Biological Bone Development Cardiac Cell Survival Cell physiology Cells Cellular Stress Complex Condition Coupled Data Development Dominant-Negative Mutation Ensure Equilibrium Event Exhibits Funding G-substrate GTP Binding GTP-Binding Proteins Glutamine Goals Guanine Nucleotides Guanosine Diphosphate Guanosine Triphosphate HL-60 Cells Hydrolysis Lead Light Link Lysine Mediating Molecular Mus Mutation NGFR Protein Natural regeneration Nerve Growth Factor Receptors Nerve Growth Factors Neurons Nucleotides Numbers Outcome PC12 Cells Pathway interactions Pheochromocytoma Play Polyamines Protein Binding Proteins Range Rattus Reaction Regulation Research Personnel Retinoblastoma Genes Retinoblastoma Protein Roentgen Rays Role Running Series Signal Pathway Signal Transduction Signaling Protein Stimulus Structure System Testing Tissues Transcriptional Activation Transferase Transglutaminases Tretinoin Up-Regulation Work aluminum fluoride amino group axon growth base cdc42 GTP-Binding Protein cell growth covalent bond cytotoxic design eIF-5A extracellular glutamine transport protein insight interest mouse Gdi2 protein mutant polymerization prevent programs protein function receptor coupling response structural biology three dimensional structure

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项目摘要

DESCRIPTION (provided by applicant): Tissue transglutaminase (TGase) is an approximately 80 kDa protein that exhibits GTP-binding and hydrolytic activities like signaling G proteins, as well as an enzymatic (transamidation) activity that catalyzes covalent linkages between glutamine residues and primary amino groups, leading to the formation of new protein-protein and protein-polyamine complexes. TGase has been implicated in a number of important biological responses including neuronal development and degeneration, as well as cellular differentiation and apoptosis. During the past funding period, we have found that TGase plays a key role in retinoic acid (RA)-induced differentiation by maintaining cell viability and protecting against apoptotic signals. Both the GTP-binding and transamidation activities of TGase, and its interactions with the retinoblastoma protein (Rb), all appear to be linked to its survival activity. We now propose that TGase-mediated cell survival may be an important outcome of a number of different extracellular stimuli and cell signaling pathways. The studies outlined in this renewal application will set out to directly test these ideas and further our understanding of the molecular mechanisms underlying TGase-mediated cell survival and the intricate regulation of this interesting protein. Three main experimental aims are proposed. 1.) Establish a role for the GTP-binding/GTP hydrolytic activities of TGase in cell survival. These studies will take advantage of our recently determined X-ray crystal structure for GDP-bound TGase to introduce mutations that perturb specific steps in the GTP-binding/GTP hydrolytic cycle and thus can serve as new dominant-active and dominant-negative TGase mutants in cellular studies. 2.) Establish a role for Rb in TGase-mediated survival. The importance of the stable binding of TGase to Rb, as well as the transamidation of Rb, for TGase-mediated cell survival and different cellular functions of Rb will be determined. 3,) Delineation of signaling pathways leading to the up-regulation of TGase expression. Here we will establish the general role that TGase activity plays in cell survival and in the regulation of apoptotic programs by examining the signaling pathways that lead from the nerve growth factor (NGF) receptor as well as from 13-amyloid to TGase expression in rat pheochromocytoma (PC12) cells. By better understanding the mechanisms of action and regulation of this important GTP-binding protein/acyl transferase, we expect to gain fundamental information regarding the balance between cell growth and differentiation versus apoptosis.

描述（由申请人提供）：组织转谷氨酰胺酶（TGASE）是一种大约80 kDa蛋白，表现出GTP结合和水解活性（如信号G蛋白），以及酶促（跨损伤）活性，催化谷氨酸残基和蛋白质蛋白质的蛋白质和蛋白质的蛋白质，可促进谷氨酰胺残基之间的共线。 TGase与许多重要的生物学反应有关，包括神经元发育和变性，以及细胞分化和凋亡。在过去的资金期间，我们发现TGASE通过维持细胞活力并预防凋亡信号来在视黄酸（RA）诱导的分化中起关键作用。 TGASE的GTP结合和跨增强活性及其与视网膜母细胞瘤蛋白（RB）的相互作用似乎都与其存活活性有关。现在，我们建议TGase介导的细胞存活可能是许多不同细胞外刺激和细胞信号通路的重要结果。在此更新应用中概述的研究将旨在直接测试这些思想，并进一步了解TGase介导的细胞存活和这种有趣蛋白质的复杂调节。提出了三个主要的实验目标。 1.）确定TGase在细胞存活中的GTP结合/GTP水解活性的作用。这些研究将利用我们最近确定的GDP结合TGase的X射线晶体结构，以引入突变，这些突变在GTP结合/GTP水解周期中扰动特定步骤，因此可以作为细胞研究中的新的主要活性和优势活性TGase突变体。 2.）在TGase介导的生存中确定RB的作用。将确定TGASE与RB的稳定结合以及RB的跨增强的重要性，即将确定TGASE介导的细胞存活和RB的不同细胞功能的重要性。 3，）信号通路的描述导致TGase表达的上调。在这里，我们将通过检查从神经生长因子（NGF）受体引起的信号传导途径，以及从13-淀粉样蛋白到TGASE在大鼠植物学细胞瘤（PC12）细胞中从13-淀粉样蛋白到TGase表达，从而确定TGASE活性在细胞存活和凋亡程序调节中发挥作用的总体作用。通过更好地了解这种重要的GTP结合蛋白/酰基转移酶的作用机理和调节的机理，我们希望获得有关细胞生长与分化与凋亡之间平衡的基本信息。