A NEW SYSTEM FOR THE STUDY OF ERYTHROPOIESIS

研究红细胞生成的新系统

• 批准号: 3238377
• 负责人: ARTHUR J SYTKOWSKI
• 金额: $ 34.19万
• 依托单位: NEW ENGLAND DEACONESS HOSPITAL
• 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 1992-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3238377
• 关键词: Rauscher's virus; Retroviridae disease; anemia; beta adrenergic agent; butyrates; cell differentiation; cell growth regulation; chromatography; congenital blood disorder; cytoskeleton; electrochemistry; electrophoresis; erythrocyte membrane; erythroleukemia; erythropoiesis; erythropoietin; gene expression; genetic library; genetic manipulation; genetic markers; hemoglobin; hemoprotein biosynthesis; hormone receptor; laboratory rabbit; membrane permeability; membrane potentials; molecular pathology; monoclonal antibody; nucleic acid probes; phosphoproteins; protein kinase; receptor; spectrin

Erythropoietin is a prime regulator of red cell development. Substantial progress has been made in studying its structure. However, very little is known about its mode of action, especially on the molecular level. The proposed research is designed to explore the action of pure, recombinant erythropoietin on the continuous erythropoietin-responsive cell line Rauscher murine erythroleukemia. We will examine the mechanism by which the signal resulting from the erythropoietin-receptor interaction is transmitted to the interior of the cell. Recent evidence indicates that erythropoietin rapidly alters the phosphorylation of one or more proteins in the erythroid cell membrane. These erythropoietin-sensitive phosphoproteins (ESPPs) may serve as second messengers for the hormone's action. This study will focus on the characterization of these ESPPs and on the elucidation of their functions and role in controlling erythroid differentiation. The ESPPs will be identified and characterized. This includes isolation and purification, protein sequence analyses, and isolating and sequencing corresponding cDNA clones. Their subcellular distribution will be examined using specific antibodies, and alterations in this distribution accompanying erythropoietin induction will be explored. The presence and distribution of these ESPPs in cells that are nonresponsive to erythropoietin will be assessed also. Radiolabeled cDNA probes will be employed to quantify changes in ESPP expression during erythropoiesis and to determine if this might serve as a controlling factor. The protein kinases involved in these phosphorylation reactions will be investigated and their relationship to erythropoietin and to its receptor will be determined. This research program represents the beginning of a systematic study of erythropoietin's action on a molecular level. The knowledge gained will be integral to our understanding of normal erythropoiesis and should prove relevant to the pathophysiology of disorders of red cell production.

红细胞生成素是红细胞发育的主要调节剂。 在研究其结构方面取得了重大进展。 但是，对其行动方式知之甚少，尤其是 在分子水平上。 拟议的研究旨在 探索纯，重组红细胞生成素对 连续红细胞生成素响应性细胞系line鼠鼠 红血球血症。 我们将研究 促红细胞生成素受体相互作用引起的信号是 传输到细胞的内部。 最近的证据表明 红细胞生成素迅速改变了一个或 红细胞膜中的更多蛋白质。 这些 促红细胞生成素敏感的磷蛋白（ESPP）可以用作 第二使激素动作的使者。 这项研究将重点 关于这些ESPP的表征和阐明 它们在控制红系分化方面的功能和作用。 ESPP将被识别和表征。 这包括 分离和纯化，蛋白质序列分析和分离 和测序相应的cDNA克隆。 他们的亚细胞 分布将使用特定抗体进行检查，并且 伴随红细胞生成素的分布的改变 将探索归纳。 这些的存在和分布 细胞中无反应性红细胞生成素的ESPP将是 也评估。 放射标记的cDNA探针将用于 量化红细胞生成期间ESPP表达的变化 确定这是否可能是控制因素。 蛋白质 这些参与这些磷酸化反应的激酶将是 调查及其与促红细胞生成素及其的关系 受体将被确定。 该研究计划代表 对促红细胞生成素对某人作用的系统研究的开始 分子水平。 获得的知识将是我们不可或缺的 了解正常的红细胞生成，应证明相关 致红细胞生产疾病的病理生理学。