JUN Kinase Signaling in the Lung

肺部的 JUN 激酶信号传导

• 批准号: 6684629
• 负责人: Glenn D. Rosen
• 金额: $ 34.74万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-04 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6684629
• 关键词: JUN kinase; apoptosis; biological signal transduction; enzyme activity; fibroblasts; guanine nucleotide binding protein; guanine nucleotide exchange factors; immunoprecipitation; lung; mitogen activated protein kinase; mitogens; nuclear factor kappa beta; oxidative stress; phosphorylation; protein protein interaction; protein signal sequence; site directed mutagenesis; tissue /cell culture; tumor necrosis factor alpha; western blottings

DESCRIPTION (provided by applicant): The small GTPase Ras proteins play an essential role in transducing extracellular signals that regulate cell growth, survival, and differentiation. Growth factors and cytokines activate Ras by recruiting proteins such as Son of Sevenless (SOS), which acts as guanine nucleotide exchange factors and converts inactive GDP-Ras to active GTP-Ras. Ras then transmits signals to downstream signaling pathways such as the mitogen-activated protein (MAP) kinase pathway. The MAPK pathway is composed of three protein kinases- Raf, MEK and MAP kinase. These kinases transduce Ras signals through a phosphorylation cascade. The MAPKs include c-Jun-NH2-terminal kinase (JNK), extracellular-regulated kinase (ERK) and p38. These MAPK regulate expression of diverse transcription factors that regulate cell growth, survival, and differentiation. From a yeast two-hybrid assay, we cloned a novel JNK-interacting protein that we call Ras Signaling Modifier (RSM). We find that growth factors and TNF-alpha induce phosphorylation of RSM leading to the association of RSM with Sos and Raf. This has two important consequences, the facilitation of Sos-mediated activation of Ras followed by Ras activation of Raf leading to an enhanced magnitude and duration of ERK activation. Oxidative stress activates the JNK pathway causing apoptosis while the ERK pathway protects cells from apoptosis and we observed that RSM protects fibroblasts from TNF-alpha-induced apoptosis. In the lung, oxidative stress has been shown to induce apoptosis of wound fibroblasts. Therefore, RSM may protect against oxidative stress injury in lung fibroblasts, which express abundant RSM, by activating the ERK pathway and inhibiting JNK activation. In addition, the ERK pathway mediates growth factor-induced proliferation and migration of fibroblasts, processes that are critical for wound healing so that RSM may augment the proliferative and migratory response to growth factors in lung fibroblasts. We view RSM as a dynamic molecular scaffolding protein in the Ras pathway. Our proposed studies will examine how RSM regulates Ras signal transduction and affects the response of lung fibroblasts to oxidative stress. We put forth three Aims: (1) Analysis of RSM in the JNK pathway; (2) Characterize the mechanism of RSM activation of the Raf/ERK pathway; and (3) Examine RSM function in lung fibroblasts. These studies will elucidate the biological function of a novel and unique regulator of Ras signal transduction.

描述（由申请人提供）：小GT3 Ras蛋白在转导调节细胞生长、存活和分化的细胞外信号中起重要作用。 生长因子和细胞因子通过募集诸如Son of Sevenless（SOS）的蛋白质来激活Ras，SOS充当鸟嘌呤核苷酸交换因子并将失活的GDP-Ras转化为活性的GTP-Ras。 Ras然后将信号传递到下游信号传导途径，如促分裂原活化蛋白（MAP）激酶途径。 MAPK通路由三种蛋白激酶组成- Raf、MEK和MAP激酶。 这些激酶通过磷酸化级联反应传递信号。 MAPK包括c-Jun-NH 2-末端激酶（JNK）、细胞外调节激酶（ERK）和p38。 这些MAPK调节调节细胞生长、存活和分化的多种转录因子的表达。 从酵母双杂交试验中，我们克隆了一种新的JNK相互作用蛋白，我们称之为Ras信号调节剂（RSM）。 我们发现生长因子和TNF-α诱导RSM的磷酸化，导致RSM与Sos和Raf的关联。 这有两个重要的后果，促进Sos介导的Ras激活，随后Ras激活Raf，导致ERK激活的幅度和持续时间增加。 氧化应激激活JNK通路引起细胞凋亡，而ERK通路保护细胞免于细胞凋亡，我们观察到RSM保护成纤维细胞免于TNF-α诱导的细胞凋亡。 在肺中，氧化应激已显示诱导伤口成纤维细胞的凋亡。 因此，RSM可能通过激活ERK通路和抑制JNK活化来保护表达丰富RSM的肺成纤维细胞免受氧化应激损伤。 此外，ERK途径介导生长因子诱导的成纤维细胞增殖和迁移，这是伤口愈合的关键过程，因此RSM可增强肺成纤维细胞对生长因子的增殖和迁移反应。 我们认为RSM作为一个动态的分子支架蛋白的Ras途径。 我们提出的研究将探讨RSM如何调节Ras信号转导和影响肺成纤维细胞对氧化应激的反应。 我们提出了三个目标：（1）分析RSM在JNK通路中的作用;（2）表征RSM激活Raf/ERK通路的机制;和（3）检查RSM在肺成纤维细胞中的功能。 这些研究将阐明Ras信号转导的一种新的和独特的调节剂的生物学功能。