GENE CONTROL AND CELL GROWTH REGULATION BY INTERFERON

干扰素的基因控制和细胞生长调节

• 批准号: 2672118
• 负责人: JAMES E DARNELL
• 金额: $ 40.04万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-01 至 2001-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2672118
• 关键词: DNA binding protein; Drosophilidae; RNase protection assay; X ray crystallography; biological signal transduction; cell growth regulation; crosslink; developmental genetics; dimer; gene expression; immunoprecipitation; interferons; intermolecular interaction; intracellular transport; mass spectrometry; phosphorylation; protein kinase; protein purification; protein structure function; protein tyrosine kinase; protein tyrosine phosphatase; proteolysis; site directed mutagenesis; transcription factor

The experiments described in this proposal deal with the action on animal cells of extracellular signalling polypeptides through a newly discovered signalling pathway, the JAKSTAT pathway. First uncovered by studying the molecular mechanism of action of interferon alpha and gamma, this pathway is now known to be the direct route by which many different cytokine and growth factors bound to their cognate receptors at the cell surface activate transcription of specific genes in the cell nucleus. This gene activation is usually transient with long lasting effects on cells due to a cascade of events dependent on the first round of gene activation. The specific experiments proposed will advance knowledge of the physical and biochemical properties of the STAT proteins, so-called because they carry out the dual function of signal transduction and activators of transcription. The first two of these proteins to be recognized, Stat1 and Stat2, will be studied intensively to determine specific amino acid contact sites for homo- and heterodimerization, a necessary step in activation, for precise amino acid DNA contact sites and for contact sites with proteins in the transcriptional machinery. Crystallographic analysis of a portion (amino acids 133-712) of Stat1 will be carried out. The mechanism of inactivation of the activated STATs, which occurs within a few minutes to a few hours will be investigated to determine whether nuclear dephosphorylation or protein turnover occurs. Among the most important biologic events concerned with signalling from the cell surface are growth control and differentiation. STAT activation functions in both events and will be studied in both contexts. IFN-alpha and gamma block entry of cells into the S phase and the IFN-alpha and gamma sensitive genes contributing to cell growth restraint will be identified. Finally, genetic and biochemical studies of development is most advanced using Drosophila and we have discovered the first Drosophila Stat molecule that will be studied for its role in developmental pathways. Because many cytokines and growth factors that are crucial in inflammation, immunity, growth regulation and normal development act through this pathway these studies have obvious implication in human disease.

本提案中描述的实验涉及动物的动作 细胞外信号传导多肽通过新发现的细胞 信号通路，jakstat途径。首先通过研究 干扰素α和伽马作用的分子机理，该途径 现在已知是许多不同的细胞因子和 与其在细胞表面的同源受体结合的生长因子 激活细胞核中特定基因的转录。这个基因 激活通常是瞬态的，由于 一连串的事件取决于第一轮基因激活。这 提出的具体实验将提高对身体和身体的了解 统计蛋白的生化特性，因为它们携带 取出信号转导和激活因子的双重功能 转录。这些蛋白质中的前两种被识别为STAT1和 STAT2将进行深入研究以确定特定的氨基酸 接触站点进行同二聚和异二聚体，这是必要的一步 激活，用于精确的氨基酸DNA接触位点和接触位点 带有转录机械中的蛋白质。晶体学分析 将执行STAT1的一部分（氨基酸133-712）。这 激活统计的灭活机制，发生在 将研究几分钟到几个小时，以确定是否 发生核去磷酸化或蛋白质更新。最重要的是 与细胞表面信号有关的重要生物事件 是生长控制和分化。 Stat激活在这两个方面都有功能 事件并将在两种情况下进行研究。 IFN-Alpha和Gamma块 将细胞进入S相以及IFN-Alpha和Gamma敏感 将确定导致细胞生长约束的基因。最后， 使用的遗传和生化研究最先进 果蝇，我们发现了第一个果蝇统计分子 将研究其在发育途径中的作用。因为很多 细胞因子和生长因子在炎症，免疫力中至关重要， 通过这一途径，增长调节和正常发展法 研究对人类疾病有明显的影响。