ERYTHROPOIETIN RECEPTOR REGULATION OF ERYTHROPOIESIS

描述（取自应用程序） 本提案的研究目标是研究分子和细胞 参与调节红细胞生成的机制， 促红细胞生成素受体（EpoR），其表达对于 红细胞发育正常。 拟议的研究包括表征 人EpoR中非启动子相关的顺式作用调控元件 基因和鉴定与该基因相互作用的反式作用因子。 调控元件，并可指导组织特异性转录控制 的基因。 将在组织中检查基因的染色质结构 培养细胞和小鼠组织，以鉴定组织和发育 特定阶段的监管要素变化。 重组dna 技术将用于克隆EpoR基因的基因组DNA片段 与其表达相关，然后进行基因转移/表达研究， 组织培养细胞以及DNA-蛋白质相互作用的研究。 另一 本研究的目的是研究细胞膜的信号转导功能， 人EpoR使用两种人红细胞生成障碍作为模型， 红细胞增多症和真性红细胞增多症。 一种新的人类EpoR的作用 引起显性遗传性红细胞增多症的突变体激活 激活的EpoR的下游靶点，如Jak2、Stat5、Shc和MAP 激酶，将进行研究，以表征信号转导异常相关 红系祖细胞的Epo高反应性。 组织培养 表达突变型和/或野生型EpoR的细胞和红系祖细胞 将在Epo dos反应测定中研究受影响个体的DNA， 片段化研究、电泳凝胶迁移率变动分析和 免疫沉淀和Western印迹实验。 在另一个模型中， 从真性红细胞增多症患者中纯化的红系祖细胞将被 研究细胞内信号分子的活化动力学 是激活的EpoR的靶点，如Jak2和Stats， 其活动过度可能与体外 对EPO的反应。本实验室研究将由 首席研究员穆拉特博士O. Arcasoy与一个教学 该计划的指导下的赞助商，伯纳德G。忘记和一 咨询委员会。 该提案将允许候选人开发 技术专长、基础知识和科学知识 独立和富有成效的职业调查所必需的方法 分子和细胞生物学及其在 血液系统疾病

DESCRIPTION (taken from application) The research goal of this proposal is to study the molecular and cellular mechanisms involved in the regulation of erythropoiesis mediated by the erythropoietin receptor (EpoR), the expression of which is essential for norma erythroid development. The proposed studies include characterization of nonpromoter-related, cis-acting regulatory elements within the human EpoR gene and identification of trans-acting factors which interact with the regulatory elements and may direct tissue-specific transcriptional control of the gene. The chromatin structure of the gene will be examined in tissue culture cells a well as mouse tissues to identify tissue-and developmental stage-specific changes in the regulatory elements. Recombinant DNA technology will be used in cloning of genomic DNA segments of the EpoR gene relevant to its expression followed by gene transfer/expression studies in tissue culture cells as well a studies of DNA-protein interactions. Another goal of this proposal is to study the signal transduction function of the human EpoR using two human disorders o erythropoiesis as models, familial erythrocytosis and polycythemia vera. The effect of a novel human EpoR mutant causing dominantly-inherited erythrocytosi on the activation of downstream targets of the activated EpoR, such as Jak2, Stat5, Shc and MAP kinase, will be studied to characterize signaling abnormalities associated with Epo hyper-responsiveness of erythroid progenitors. Tissue culture cells expressing the mutant and/or wild type EpoRs and erythroid progenitors from affected individuals will be studied in Epo dos response assays, DNA fragmentation studies, electrophoretic gel mobility shift assays and immunoprecipitation and Western blotting experiments. In another model, purified erythroid progenitors from patients with polycythemia vera wil be studied for the activation kinetics of intracellular signaling molecules that are targets for the activated EpoR, such as Jak2 and Stats, hyperactivity of which may be associated with increased in vitro responsiveness to Epo. This laboratory research will be performed by the principal investigator Dr. Murat O. Arcasoy in conjunction with a didactic program under the guidance of a sponsor, Dr. Bernard G. Forget and an advisory committee. This proposal will allow the candidate to develop the technical expertise, fundamental knowledge and intellectual scientific approach necessary for an independent and productive career in investigative molecular and cellular biology and its application to pathogenesis of hematologic disorders.