REGULATION OF ERYTHROID TRANSCRIPTION

红细胞转录的调节

• 批准号: 2227457
• 负责人: NANCY CATHERINE ANDREWS
• 金额: $ 10.52万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-12-01 至 1998-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2227457
• 关键词: DNA binding protein; antibody; cell differentiation; chromatin; complementary DNA; dimer; erythroid stem cell; erythropoiesis; gel mobility shift assay; genetic enhancer element; genetic models; genetic regulation; genetic transcription; hemoglobin; laboratory rabbit; molecular cloning; nucleic acid probes; posttranslational modifications; protein sequence; protein structure function; site directed mutagenesis; transcription factor; western blottings

Differentiation of pluripotential hematopoietic stem cells into mature blood components requires lineage-specific regulation of gene expression. The general aim of this project is to understand transcriptional regulation of erythropoiesis, using globin gene expression as a model. Mammalian alpha-like and beta-like globin genes are organized in cluster, each of which is associated with an upstream regulatory element termed the locus control region (LCR). LCRs enhancers. They are made up of three DNA motifs which bind nuclear regulatory proteins: CACCC motifs binding ubiquitous transcription factors. GATA motifs binding erythroid factor GATA-1 and AP-1 like motifs binding erythroid factor NF-E2. Of these proteins, NF-E2 acts as the primary regulator of LCR enhancer function. It appears to be a dimer of a 45kD tissue-specific basic- leucine zipper (bZIP) protein (p45 NF-E2), and a smaller protein has remained elusive. This investigation is aimed at understanding how NF-E2 mediates LCR enhancer activity. The first step will be to clone p18, using peptide sequences derived from purified protein. Although not essential for further studies, cloning of p 18 will facilitate investigation of the relationship between NF-E3 protein structure and function. Assays of various aspects of LCR activity will be developed using defined components, an employed to examine the functional effects of deliberate mutations of NF-E3. Finally, interactions of p45 NF-E2 and p18 with other partners will be explored, to understand regulation at the level of differential dimerization. Elucidation of the role of NF-E2 in globin gene expression and erythropoiesis will contribute to our overall understanding of the genetic and molecular mechanisms responsible for controlling hematopoietic commitment and differentiation. At the same time, progress will be made towards understanding disorders of erythropoiesis, including thalassemias, congenital anemias, and red cell hypoplasia from HIV infection.

多能造血干细胞的分化成熟 血液成分需要特定于基因表达的谱系调节。 该项目的总体目的是了解转录 使用球蛋白基因表达作为模型的红细胞生成的调节。 哺乳动物α样和类似β的球蛋白基因在簇中组织， 每一个都与所谓的上游调节元件有关 基因座控制区（LCR）。 LCRS增强剂。 他们由 结合核调节蛋白的三个DNA基序：CACCC基序 结合无处不在的转录因子。 GATA主题结合红细胞 因子GATA-1和AP-1像结合红系因子NF-E2的基序。 的 这些蛋白质NF-E2充当LCR增强剂的主要调节剂 功能。 它似乎是45kD组织特异性基本的二聚体 - 亮氨酸拉链（BZIP）蛋白（p45 NF-E2），较小的蛋白质具有 仍然难以捉摸。 这项调查旨在了解NF-E2如何 介导LCR增强剂活性。 第一步是克隆P18， 使用源自纯化蛋白的肽序列。 虽然不是 对于进一步的研究至关重要，P 18的克隆将有助于 研究NF-E3蛋白结构与 功能。 LCR活动的各个方面的测定将开发 使用定义的组件，用于检查功能效应 NF-E3的故意突变。 最后，P45 NF-E2和 将探索与其他合作伙伴的P18，以了解 差异化水平。 阐明NF-E2在 Globin基因表达和红细胞生成将有助于我们的整体 了解负责的遗传和分子机制 控制造血承诺和分化。 同样 时间，将取得进步来理解的疾病 红细胞生成，包括thalassemias，先天性贫血和红细胞 HIV感染发育不全。