REGULATION OF PROTEIN SYNTHESIS AND ERYTHROPOIESIS

这项研究的长期目标是阐明 正常和异常血液病患者血红蛋白合成的调节 条件；重点放在对 红系细胞合成珠蛋白和其他蛋白质的研究 2α-激酶及其与红系细胞的关系 发展和成熟。具体目的包括澄清： 血红素调控的双链RNA的结构与功能关系 激活的eIF-2α激酶在人类血液病中的作用 以血红素缺乏为特征的。这项研究与真核生物有关。 一般情况下，尤其是对于理解 发生血红素缺乏的人类贫血：缺铁性贫血， 世界上最流行的疾病，铅中毒引起的贫血，以及 铁粒母细胞性贫血。实验设计和实验方法包括： 兔网织红细胞蛋白质原位合成及其功能的研究 这些细胞的裂解产物和体外成分；纯化 血红素调节的eIF-2α激酶；血红素和 EIF-2α激酶上的其他金属卟啉；以及 EIF-2α激酶的cDNA及其氨基酸的测定 序列及其三磷酸腺苷结合位点；多克隆和 抗eIF-2α蛋白激酶的单抗及其检测 红系组织中eIF-2α激酶表达的免疫荧光研究 正常人和血红素中不同发育阶段的细胞- 虚证临床障碍。

The long-term objectives of this research are the elucidation of the regulation of hemoglobin synthesis in normal and abnormal hematologic conditions; emphasis is placed on the translational control of the synthesis of globin and other proteins in erythroid cells by heme and eIF- 2alpha kinases and on the relationship of this control to erythroid cell development and maturation. The specific aims include elucidation of: the structure-function relations of heme-regulated and double-stranded RNA activated eIF-2alpha kinase in the human hematologic disorders characterized by heme-deficiency. This research is relevant to eukaryotic cells generally and specifically it is significant for understanding the human anemias in which heme-deficiency occurs: iron-lack anemia, one of the most prevalent world-wide disorders, the anemia of lead poisoning, and the sideroblastic anemias. The experimental design and methods involve: the study of protein synthesis in situ in rabbit reticulocytes and their lysates and in vitro in components of these cells; the purification of the heme-regulated eIF-2alpha kinase; the study of the effects of heme and other metalloporphyrins on the eIF-2alpha kinase; and the cloning of the cDNAs of the eIF-2alpha kinases, the determination of their amino acid sequences and their ATP binding sites; the preparation of polyclonal and monoclonal antibodies to the eIF-2alpha kinases and the determination by immunofluorescence of the presence of the eIF-2alpha kinases in erythroid cells at different stages of development in normal man and in the heme- deficiency clinical disorders.