DEVELOPMENTAL BIOLOGY OF HUMAN ERYTHROPOIESIS

建议的计划(改编自申请人的摘要)长期 本应用程序的目标是在分子水平上理解 红细胞膜蛋白在细胞水平上的作用 生长因子受体、信号转导蛋白与转录 红细胞生成过程中的因素。洛迪什博士将专注于最常见的 IL-3/GM-CSF/IL-5受体和促红细胞生成素受体链， 分别进行了分析。它们在红系分化中的作用将是 通过使用嵌合受体结构、受体 细胞质结构域融合蛋白鉴定转导蛋白和 它们的结合部位，显性负/正信号突变体 蛋白质，如STAT和MAP激酶，评估它们在 在细胞系、胎肝和胚胎干细胞中体外培养，以及在 体内转基因和靶向基因置换的敲入小鼠。Dr。 Zon将通过识别新的激酶来补充这些研究 不同的MAP激动级联反应及其在细胞周期中的作用 红系细胞的发育。该项目还将提供一座桥梁 为Orkin博士的项目提供了定义三个角色的方法 不同的MAP激酶在转导来自IL-3和 促红细胞生成素受体，并在激活红系特异性 转录因子。奥尔金博士的项目将专注于 GATA-1在红系细胞中作用的结构-功能研究 GATA因子依赖的末端关键靶基因的鉴定 红系发育。Lux博士将研究Ankyrin的功能 红系细胞通过利用靶向基因干扰和基因 胚胎干细胞的替代策略。通过这些手段，我们希望了解 更全面地了解红细胞生成过程并获得分子洞察力 转化为功能衰竭、生产异常或增殖增加的紊乱 红系细胞。

PROPOSED PROGRAM (adapted from applicant's abstract) The long-term objectives of this application are to understand at a molecular and cellular level the roles of erythroid membrane proteins, hematopoietic growth factor receptors, signal transduction protein, and transcription factors during erythropoiesis. Dr. Lodish will focus on the b common chain of the IL-3/GM-CSF/IL-5 receptor and the erythropoietin receptor, respectively. Their role in erythroid differentiation will be investigated through the use of chimeric receptor constructs, receptor cytoplasmic domain fusion proteins to identify transduction proteins and their binding sites, dominant negative/positive mutants of signalling proteins such as stat and MAP kinases, assessing their function both in vitro in cell lines and fetal liver and embryonal stem (ES) cells, and in vivo in transgenic and targeted gene replacement of knock-in mice. Dr. Zon will compliment these studies by identifying novel kinases of different MAP kinase cascades and assessing their role during the development of erythroid cells. This project will also provide a bridge to Dr. Orkin's project by providing the means to define the role of three different MAP kinase cascades in transducing signals from the IL-3 and erythropoietin receptors, and in activating erythroid specific transcription factors. Dr. Orkin's project will focus on the structure-function studies of the role of GATA-1 in erythroid cells and identification of GATA factor dependent target genes critical for terminal erythroid development. Dr. Lux will study the function of ankyrin in erythroid cells through the use of targeted gene disruption and gene replacement strategies in ES cells. By these means we hope to understand more fully the process of erythropoiesis and obtain molecular insights into disorders of failure, abnormal production, or increased proliferation of erythroid cells.