Regulation of signaling via the lkB-NF-kB interaction

通过 lkB-NF-kB 相互作用调节信号传导

• 批准号: 7249466
• 负责人: Alexander Hoffmann
• 金额: $ 22.25万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7249466
• 关键词: 26S proteasome; Binding; Biochemistry; Cell Death; Cell Line; Cells; Characteristics; Chronic; Clinical Treatment; Computer Simulation; Disease; Equilibrium; Gene Expression; Gene Targeting; Genetic; Goals; Half-Life; Health; Human; I Kappa B-Alpha; Inflammatory; Interphase Cell; Knock-out; Knockout Mice; Lentivirus Vector; Life; Malignant Neoplasms; Measures; Mediating; Mus; Mutate; Mutation; NF-kappa B; Noise; Performance; Phosphotransferases; Physiological; Physiology; Principal Investigator; Protein Biosynthesis; Protein Isoforms; Proteins; Proteolysis; Publishing; Rate; Regulation; Relative (related person); Resistance; Role; Signal Pathway; Signal Transduction; Signaling Protein; Small Interfering RNA; Specificity; Stimulus; Subfamily lentivirinae; System; Transgenes; Work; casein kinase II; dimer; genome-wide analysis; human disease; in vivo; inhibitor/antagonist; knockout gene; lymphocyte proliferation; mutant; neglect; promoter; protein degradation; reconstitution; research study; response; small molecule; transcription factor; tumorigenesis; ultraviolet irradiation

DESCRIPTION (provided by applicant): The transcription factor NF-kappaB is critical in inflammatory gene expression, lymphocyte proliferation and cell death/survival decisions. Its activity is controlled by three inhibitory proteins, iKappaBalpha -beta -epsilon, whose synthesis and protein half-life is tightly regulated. Stimulus-induced degradation of IkappaB proteins associated with NFkappaB studied in great detail. The goals of the work proposed here are to characterize and contrast half-life regulation of NF-kappaB-bound and "free" IkappaB proteins in resting cells and those responding to inflammatory stimuli and UV irradiation. The mechanisms and potential signaling pathways that determine these degradation rates will be characterized, and importantly, the role of the interaction with NF-kappaB in determining half-regulation will be examined. These results will serve to refine a computational model of NF-kappaB signaling significantly, that will allow us to examine the role of regulating these previously neglected degradation rates in NF-kapppaB signaling. First, we will focus on the equilibrium state in resting cells: Robustness of the signaling system vis-a-vis transient perturbations ("noise") or long-term perturbations due to chronic changes in expression levels of the signaling components for example. Such perturbations may be caused by genetic changes that may be associated with inflammatory disease or tumorigenesis. Next, we will focus on the performance of the IkappaB/NF-kappaB signaling system during signal transduction, especially in response to weak or transient stimuli. Computational predictions will be examined experimentally using mutant cell lines generated from knockout mice and by lentivirus-mediated reconstitution of these with IkappaB mutant molecules. We will identify NF-kappaB target genes whose expression is sensitive to the regulation of IkappaB degradation rates and thereby demonstrate the specificity and physiological relevance of these mechanisms in human health and disease.

描述（由申请人提供）：转录因子NF-kappab在炎症基因表达，淋巴细胞增殖和细胞死亡/生存决策中至关重要。它的活性由三种抑制蛋白Ikappabalpha -beta -epsilon控制，其合成和蛋白质半衰期受到严格调节。刺激引起的与NFKappab相关的Ikappab蛋白的降解非常详细地研究。这里提出的工作的目标是表征和对比静止细胞中NF-kappab结合和“自由” ikappab蛋白的半衰期调节，以及对炎症刺激和紫外线照射的反应的人。确定这些降解率的机制和潜在信号通路将被表征，重要的是，将检查与NF-kappab在确定半调节中的相互作用的作用。这些结果将大大改善NF-kappab信号传导的计算模型，这将使​​我们能够检查调节这些先前被忽略的降解速率在NF-kapppab信号传导中的作用。首先，我们将重点关注静息细胞中的平衡状态：信号系统的鲁棒性相对于瞬态扰动（“噪声”）或由于信号传导成分表达水平的长期变化而导致的长期扰动。这种扰动可能是由遗传变化引起的，可能与炎症性疾病或肿瘤发生有关。接下来，我们将重点介绍信号转导期间Ikappab/NF-kappab信号系统的性能，尤其是响应弱或瞬态刺激。将使用由基因敲除小鼠产生的突变细胞系以及慢病毒介导的重构与Ikappab突变分子对这些重构进行实验研究。 我们将确定NF-kappab靶基因的表达对IKAPPAB降解率的调节敏感，从而证明了这些机制在人类健康和疾病中的特异性和生理相关性。