HEMATOPOIETIC RING FINGER 1 (HERF1) IN ERYTHROPOIESIS

红细胞生成中的造血环指 1 (HERF1)

• 批准号: 6501111
• 负责人: JAMES R DOWNING
• 金额: $ 12.63万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6501111
• 关键词: cell line; erythroid stem cell; erythropoiesis; gene expression; laboratory mouse; protein structure function; sickle cell anemia

This proposal focuses on defining at a mechanistic level, the role in erythropoiesis of a newly discovered gene, HERF1, that we cloned as a downstream target of the AML1/CBFbeta transcription factor complex. The HERF1 protein contains an N-terminal RING-finger domain followed by a single B-box motif, a leucine coiled-coil domain, and a C-terminal rfp homologous region. Expression of HERF1 during embryogenesis coincides with the appearance of definitive erythropoiesis and, in adult mice, is restricted to the erythroid lineage, increasing over 40-fold during terminal differentiation. Importantly, we have demonstrated that inhibition of HERF1 expression blocks terminal erythroid differentiation, whereas over-expression of HERF1 in erythroid cells induces beta-major globin expression and erythroid differentiation. Taken together, these results suggest that HERFF1 plays an essential role in the development of mature erythroid cells. The studies in this project seek to elucidate, at a mechanistic level, the role of HERF1 in the linage commitment and maturation of erythroid progenitors. Our working hypothesis is that HERF1 functions as a part of a multiprotein complex to regulate cellular pathways required for the development and terminal differentiation of erythroid cells. Experiments are proposed in specific aim 1 to examine the biologic role of HERF1 in erythroid development through generation of both HERF1-deficient ES cells and mice, as well as murine systems with enforced HERF1 expression. In experiments outlined in specific aim 2, we will investigate the biochemical mechanism of action of HERF1 to gain insight into its mechanistic role in erythroid cell development. Together, these studies should provide important information on critical regulatory pathways required for the normal development of cells of the erythroid lineage. This fundamental knowledge is likely to expand our abilities to rationally manipulate erythroid lineage functions, potentially for therapeutic purposes in sickle cell anemia

该建议的重点是在机制水平上定义新发现的基因HERF 1在红细胞生成中的作用，我们克隆了该基因作为AML 1/CBFbeta转录因子复合物的下游靶点。HERF 1蛋白含有一个N-末端环指结构域，随后是一个单一的B-盒基序，一个亮氨酸卷曲螺旋结构域和一个C-末端rfp同源区。在胚胎发生过程中，HERF 1的表达与定形红细胞生成的出现相一致，并且在成年小鼠中，仅限于红细胞系，在终末分化过程中增加超过40倍。重要的是，我们已经证明，抑制HERF 1表达阻断终末红系分化，而在红系细胞中HERF 1的过表达诱导β-主要珠蛋白表达和红系分化。 总而言之，这些结果表明HERFF 1在成熟红系细胞的发育中发挥着重要作用。本项目的研究旨在从机制层面阐明HERF 1在红系祖细胞谱系定型和成熟中的作用。我们的工作假设是HERF 1作为多蛋白复合物的一部分发挥作用，调节红系细胞发育和终末分化所需的细胞途径。在具体目标1中提出了实验，以通过产生HERF 1缺陷型ES细胞和小鼠以及具有增强的HERF 1表达的鼠系统来检查HERF 1在红系发育中的生物学作用。在具体目标2中概述的实验中，我们将研究HERF 1作用的生化机制，以深入了解其在红系细胞发育中的机制作用。总之，这些研究应该提供重要的信息，红系细胞正常发育所需的关键调控途径。这一基础知识很可能扩展我们合理操纵红细胞谱系功能的能力，可能用于镰状细胞贫血的治疗目的