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

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 的蛋白质，具有 GTP 结合和水解活性（如信号 G 蛋白），以及催化谷氨酰胺残基和伯氨基之间共价连接的酶促（转酰胺基）活性基团，导致新的蛋白质-蛋白质和蛋白质-多胺复合物的形成。 TGase 涉及许多重要的生物反应，包括神经元发育和变性，以及细胞分化和凋亡。在过去的资助期间，我们发现 TGase 通过维持细胞活力和防止细胞凋亡信号在视黄酸 (RA) 诱导的分化中发挥关键作用。 TGase 的 GTP 结合和转酰胺基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生存活性有关。我们现在提出，TGase 介导的细胞存活可能是许多不同的细胞外刺激和细胞信号传导途径的重要结果。本更新申请中概述的研究将直接测试这些想法，并进一步了解 TGase 介导的细胞存活的分子机制以及这种有趣蛋白质的复杂调节。提出了三个主要实验目标。 1.) 确定 TGase 的 GTP 结合/GTP 水解活性在细胞存活中的作用。这些研究将利用我们最近确定的 GDP 结合 TGase 的 X 射线晶体结构来引入扰乱 GTP 结合/GTP 水解循环中特定步骤的突变，从而可以作为新的显性活性和显性失活 TGase 突变体在细胞研究中。 2.) 确定 Rb 在 TGase 介导的生存中的作用。将确定 TGase 与 Rb 的稳定结合以及 Rb 的转酰胺基化对于 TGase 介导的细胞存活和 Rb 的不同细胞功能的重要性。 3,) 描绘导致 TGase 表达上调的信号通路。在这里，我们将通过检查从神经生长因子 (NGF) 受体以及从 13-淀粉样蛋白到大鼠嗜铬细胞瘤中 TGase 表达的信号传导途径，确定 TGase 活性在细胞存活和细胞凋亡程序调节中的一般作用。 (PC12) 细胞。通过更好地了解这种重要的 GTP 结合蛋白/酰基转移酶的作用和调节机制，我们期望获得有关细胞生长和分化与凋亡之间平衡的基本信息。