REGULATION OF GLOBIN GENE EXPRESSION

球蛋白基因表达的调控

• 批准号: 6456254
• 负责人: KATHY ANDERSON
• 金额: $ 22.86万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6456254
• 关键词: DNA footprinting; developmental genetics; gene deletion mutation; gene expression; genetic regulatory element; globin; hemoglobin; hemoglobin Ss; hemoprotein biosynthesis; hemoprotein structure; laboratory mouse; nucleic acid sequence; sickle cell anemia; site directed mutagenesis; tissue /cell culture; transcription factor

The long term goal of our research is to define the tissue and developmental-specific regulation of the globin genes and to eventually understand erythroid cell development. An understanding of the cellular and molecular basis of the regulated expression of adult beta hemoglobin is important for two reasons. First, it will provide detailed knowledge of how genes are controlled during development and regulated in general, and second and equally important, it should provide the basis for exploring new approaches to the treatment of certain hemoglobinopathies. Clearly this would be helpful in sickle cell anemia, where an understanding at the molecular level of the mechanisms governing the switch from fetal to adult hemoglobin could present new avenues for preventing or reversing this switch. In keeping with this goal, the specific aims of our studies are as follows: 1. Characterization of a newly identified cis element in the distal 5' flanking sequence of the human beta globin gene. Our recent studies have identified a new cis element involved in the expression of the human globin gene and we are now characterizing this element and the transacting factors which interact with it. This element is particularly noteworthy as it may form a link between the beta globin gene and its locus activating region, i.e. HS2. Both deletion and site-directed mutagenesis will be used along with footprinting and oligonucleotide competition studies to localize the nucleotide binding site. Efforts to purify the factor(s) binding to this site will include the use of the yeast one-hybrid system as well as conventional approaches. 2. Regulation of the EKLF gene. We have also initiated studies on the regulation of the EKLF gene which codes for a transcription factor required for beta globin gene expression and has been implicated as one of the key elements necessary for the fetal-to-adult switch in globin gene expression. We plan to define the cis elements and transacting factors responsible for the tissue specific expression of the EKLF gene. Together, these studies will provide new insights into mechanisms of erythroid-specific gene regulation and the biology of hematopoietic systems.

我们研究的长期目标是定义组织和 球蛋白基因的发育特异性调节并最终 了解红细胞的发育。对细胞的理解 成人β血红蛋白的调节表达的分子基础 很重要，原因有两个。首先，它将提供有关的详细知识 基因在开发和一般监管过程中如何控制基因，以及 第二和同样重要的是，它应该为探索提供基础 治疗某些血红蛋白病的新方法。清楚地 这对镰状细胞贫血有帮助，在这里了解 管理从胎儿到成人转换的机制的分子水平 血红蛋白可以提出预防或逆转这一新途径 转变。为了与这个目标保持一致，我们研究的具体目的是 如下：1。在该中新识别的CIS元素的表征 人β球蛋白基因的远端5'侧翼序列。我们最近 研究已经确定了参与表达的新的顺式元素 人类球蛋白基因和我们现在正在描述这个元素和 与之相互作用的交易因子。这个元素特别是 值得注意的是，它可能形成Beta Globin基因与其之间的联系 基因座激活区域，即HS2。删除和站点定向 诱变将与足迹和寡核苷酸一起使用 竞争研究以定位核苷酸结合位点。努力 净化与此站点的因素结合的因素将包括使用 酵母单杂交系统以及常规方法。 2。法规 eklf基因。我们还开始了有关调节的研究 EKLF基因编码β球蛋白所需的转录因子 基因表达，并被牵涉到关键要素之一 Globin基因表达中胎儿向成年开关所必需的。我们计划 定义负责的顺式元素和交易因素 EKLF基因的组织特异性表达。这些研究将在一起 提供有关红斑特异性基因调节机制的新见解 以及造血系统的生物学。