NEW SYSTEM FOR THE STUDY OF ERYTHROPOIESIS

研究红细胞生成的新系统

• 批准号: 2140693
• 负责人: ARTHUR J SYTKOWSKI
• 金额: $ 28.18万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-08-01 至 1997-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2140693
• 关键词: antisense nucleic acid; biological signal transduction; cell differentiation; cell growth regulation; cytoplasm; dimethylsulfoxide; enzyme mechanism; erythrocyte membrane; erythroid stem cell; erythropoiesis; erythropoietin; gene expression; laboratory mouse; molecular cloning; okadaic acid; phosphoprotein phosphatase; phosphorylation; protein kinase C; protein sequence; protooncogene; receptor binding; regulatory gene; transfection; western blottings

The interaction of hematopoietic growth factors with their cognate receptors results in the activation of one or more signal transduction pathways leading to changes in the expression of genes that regulate cell growth and differentiation. This proposal is directed toward the elucidation of those signaling elements triggered by the hormone erythropoietin and toward the characterization of the roles played by the early response proto-oncogenes c-myc and c-myb. The individual phosphoprotein involved in membrane and cytosolic signaling will be studied. Selective proteins will be cloned and sequenced and their intracellular concentrations will be modulated with antisense oligodeoxynucleotides and by transfection with constitutively expressed clones. Protein kinase C subclasses will be investigated. Erythroid cell subclasses will be quantified and their expression will be manipulated experimentally permitting the identification of subclass specific protein phosphorylations. The role of protein phosphatases 1 and 2A in erythropoietin signaling will be evaluated using the phosphatase inhibitor okadaic acid. A detailed study and comparison of erythropoietin and dimethyl sulfoxide signals to the proto-oncogene c-myc will be undertaken. Specifically, a comparison of the membrane and cytosolic phosphorylation events utilized by these two inducers of erythroleukemia cell differentiation will be made. The molecular mechanism of erythropoietin's signal to c-myb will be studied. The hypothesis that the signal to c-myb is mediated by a phosphatase dependent reaction operating on proteins of fos/jun (AP-1) complex will be evaluated. The results of these investigations will be directly relevant to our understanding of the control of red blood cell production and may provide insights into the development of new therapies for disorders of hematopoiesis.

造血生长因子与同源的相互作用 受体导致一个或多个信号转导的激活 导致调节细胞基因表达的变化的途径 生长与分化。 该提议针对 阐明由激素触发的那些信号元素 促红细胞生成素，并朝着扮演角色的特征 早期反应原始基因C-MYC和C-MYB。 个人 参与膜和胞质信号传导的磷蛋白将是 研究。 选择性蛋白将被克隆和测序，它们 细胞内浓度将通过反义调节 寡脱氧核苷酸和通过组成型表达的转染 克隆。 将研究蛋白激酶C亚类。 红细胞 细胞亚类将被量化，它们的表达将是 操纵实验允许鉴定子类 特定的蛋白质磷酸化。 蛋白质磷酸酶的作用1 和2a在红细胞生成素信号中将使用 磷酸酶抑制剂冈田酸。 详细的研究和比较 促红细胞生成素和二甲基亚氧化二甲基亚氧化物信号到原始癌基因C-Myc 将进行。 具体而言，膜和 这两个诱导剂的胞质磷酸化事件 将进行红血球血症细胞分化。 分子 将研究促红细胞生成素对C-MYB信号的机制。 这 假设C-MYB的信号是由磷酸酶介导的 在FOS/JUN（AP-1）复合物的蛋白质上运行的依赖反应将 进行评估。 这些调查的结果将直接 与我们对红细胞产生的控制的理解有关 并可能提供有关开发新疗法的见解 造血的疾病。