Regulating IRS-proteins by Ser/Thr Phosphorylation

通过 Ser/Thr 磷酸化调节 IRS 蛋白

• 批准号: 7414864
• 负责人: Morris F. White
• 金额: $ 32.16万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-07-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7414864
• 关键词: Acute; Alleles; Antibody Formation; Build-it; Cells; Chronic; Complex; Data; Diabetes Mellitus; Docking; Funding; Genes; Genetic Polymorphism; Goals; Homologous Gene; Human; IRS1 gene; IRS2 gene; Inflammation; Insulin; Insulin Resistance; JUN kinase; Libraries; Life; Link; Literature; Mediating; Metabolic; Metabolic Diseases; Metabolic stress; Metabolism; Molecular; Mus; N-terminal; Non-Insulin-Dependent Diabetes Mellitus; Nutrient; Pancreas; Phospho-Specific Antibodies; Phosphorylation; Phosphorylation Site; Phosphotransferases; Play; Process; Production; Proteins; Publications; Regulation; Risk; Rodent; Role; Serine/Threonine Phosphorylation; Signal Pathway; Signal Transduction; Signaling Protein; Site; Sorting - Cell Movement; Source; Stream; Tissues; Tyrosine Phosphorylation; United States National Institutes of Health; Validation; cell growth; cytokine; human IRS2 protein; insulin receptor substrate 1 protein; insulin receptor tyrosine kinase; insulin sensitivity; insulin signaling; link protein; mutant; prototype; response

DESCRIPTION (provided by applicant): This application entitled "Regulating IRS-proteins by Ser/Thr Phosphorylation" is the amened competing renewal ofDK38712-16. IRS1 is a prototype docking protein that links the activated insulin receptor tyrosine kinase to down-stream signaling pathways. Tyrosine phosphorylation of IRS 1, and its homolog IRS2, activates the PI3K and ras/MAPK cascades, which promote cellular growth, survival and metabolism. Nutrient excess, acute and chronic inflammation, or proinflammatory cytokines regulate IRS-protein signaling through multisite Ser/Thr-phosphorylation. During the previous funding period, we revealed the inhibitory role of Ser307 phosphorylation in rodent IRS1 (Ser312 in human IRS1), and showed the utility of polyclonal phosphospecific antibodies to study these processes in murine and human cells and tissues. Phosphorylation of Ser307 is mediated, at least in part, by the association of the activated NH2-terminal JUN kinase (JNK) with IRS1, linking insulin signaling to the inhibitory effects of metabolic stress and proinflammatory cytokines. IRS1and IRS2 each contain at least 40 Ser/Thr-phosphorylation sites (probably more), but how they interact to regulate insulin action in various tissues under acute and chronic metabolic stress is important to resolve. Here, we propose 5 Specific Aims to reveal how Ser/Thr-phosphorylation of Irs1 and Irs2 modulates insulin sensitivity and contirbutes to life threatening metabolic diseases that can progress to diabetes. 1. Prepare and validate a comprehensive mAb library against observed S/T-phosphorylation sites in Irs1 and Irs2 (apS/TmAbIrs1 and apS/TmAbIrs2 libraries). Support is requested in year 1 to generate the library against Irs1 (apS/TmAbIrs1); the apS/TmAbIrs2 library already underway and supported by other sources. 2. Utilize the pS/TmAblrs1 and pS/TmAbIrs2 libraries to decode the S/T-phosphorylation that occurs on Irs1 and Irs2 during metabolic stress. 3. Investigate the regulatory function of Ser307 in Irs1 using mice in which the Irs1 alleles are replaced with mutant A307Irs1 alleles. 4. Determine whether Ser302 in Irsl plays a positive or negative regulatory role upon Irsl signaling in murine tissues. 5. Establish the role of the Jnk-directed "kinase interaction motif (KIM) in Irsl. This proposal is important because dysregulated insulin signaling contributes to life threatening metabolic disease that progresses to type 2 diabetes when pancreatic b-cells fail to secrete sufficient insulin quickly enough to compensate for insulin resistance. Ser/Thr-phosphorylation of Irs 1 and Irs2 is common cause of insulin resistance. The production and validation of the apS/TmAbIrs1 and apS/TmAbIrs2 libraries to detect and quantify the phosphorylation sites provides a rational experimental platform to decoding the complex regulatory mechanism that controls the insulin response throughout life.

描述（由申请人提供）：此申请名为“通过Ser/Thr磷酸化调节IRS蛋白质”是AMENED竞争的续订DK38712-16。 IRS1是一种原型对接蛋白，将活化的胰岛素受体酪氨酸激酶与下游信号通路联系起来。 IRS 1及其同源性IRS2的酪氨酸磷酸化激活了PI3K和RAS/MAPK级联反应，从而促进细胞生长，生存和代谢。营养过量，急性和慢性炎症或促炎细胞因子通过多站点SER/THR磷酸化调节IRS-蛋白质信号。在上一个资金期间，我们揭示了Ser307磷酸化在啮齿动物IRS1中的抑制作用（人类IRS1中的Ser312），并显示了多克隆磷酸特异性抗体在鼠类和人类细胞和人类细胞和组织中研究这些过程的实用性。 Ser307的磷酸化至少部分是由活化的NH2-末端JUN激酶（JNK）与IRS1的关联介导的，将胰岛素信号与代谢胁迫和促炎细胞因子的抑制作用联系起来。 IRS1和IRS2每个都包含至少40个Ser/Thr磷酸化位点（可能更多），但是它们如何相互作用以调节急性和慢性代谢应激下各种组织中的胰岛素作用对于解决方案很重要。在这里，我们提出了5个特定旨在揭示IRS1和IRS2的Ser/Thr-磷酸化如何调节胰岛素敏感性和对威胁生命的代谢性疾病的敏感性，从而可以发展为糖尿病。 1。准备并验证IRS1和IRS2（APS/TMABIRS1和APS/TMABIRS2库）中观察到的S/T磷酸化位点的综合MAB文库。在第1年要求支持IRS1（APS/TMABIRS1）的库； APS/TMABIRS2库已经在进行中，并得到了其他来源的支持。 2。利用PS/TMABLRS1和PS/TMABIRS2库来解码代谢应力期间IRS1和IRS2上发生的S/T磷酸化。 3。使用小鼠使用IRS1等位基因代替突变体A307IRS1等位基因的小鼠研究Ser307的调节功能。 4。确定IRSL中的Ser302在鼠组织中IRSL信号传导上是否扮演正调控作用。 5。建立IRSL中JNK定向的“激酶相互作用基序（KIM）的作用。该提案很重要，因为失调的胰岛素信号失调会导致威胁生命的代谢疾病，该疾病会导致2型糖尿病的发展，而当胰腺B-cells不足以使1次胰岛素足够弥补的胰岛素及其胰岛素的胰岛素共同促进胰岛素的共同点。胰岛素抵抗。