REGULATION OF PROTEIN SYNTHESIS AND ERYTHROPOIESIS

本研究的长期目标是阐明 正常和异常血液系统中血红蛋白合成的调节 条件;重点放在平移控制的 血红素和eIF在红系细胞中合成珠蛋白和其他蛋白质- 2 α激酶及其与红系细胞的关系 发展和成熟。 具体目标包括阐明： 血红素调节RNA和双链RNA结构与功能关系 人类血液系统疾病中的活化eIF-2 α激酶 以血红素缺乏为特征。 这项研究与真核生物有关。 细胞一般和具体来说，这是重要的理解， 血红素缺乏的人类贫血;缺铁性贫血， 最普遍的世界范围内的疾病，贫血的铅中毒，和 铁粒幼细胞性贫血 实验设计和方法涉及： 兔网织红细胞蛋白质合成原位研究 裂解物和体外这些细胞的组分; 血红素调节的eIF-2 α激酶;血红素和 其他金属卟啉对eIF-2 α激酶的作用; eIF-2 α激酶的cDNA，其氨基酸的测定 序列及其ATP结合位点;制备多克隆和 抗eIF-2 α激酶的单克隆抗体， 红系细胞中eIF-2 α激酶存在的免疫荧光 正常人和血红素中不同发育阶段的细胞， 缺乏症临床病症。

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.