GLOBIN SWITCHING AND ERYTHROID DIFFERENTIATION

珠蛋白转换和红细胞分化

• 批准号: 2138522
• 负责人: THALIA STAMATOYANNOPOULOS
• 金额: $ 39.67万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-06-01 至 1995-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2138522
• 关键词: cell differentiation; cell growth regulation; erythroid stem cell; erythroleukemia; gene deletion mutation; gene expression; gene rearrangement; genetic library; genetic mapping; genetic recombination; genetic transcription; hematopoiesis; hemoglobin; hemoglobin F; hemoprotein biosynthesis; human genetic material tag; human tissue; mutant; neoplastic cell; nucleic acid biosynthesis; subtraction hybridization; thalassemia; transcription factor

The long term objective of this grant proposal is to delineate the control of the switching from fetal to adult globin formation during human development. To accomplish this we will apply a somatic cell hybridization approach we have developed which allows us to do observations on the expression of normal, mutant or engineered globin genes in their normal chromosomal location. Using this system, we will investigate first, the mechanism of gamma to beta switching during ontogeny. We will attempt to obtain definitive evidence that globin gene switching is controlled in trans; we will perform studies aiming to detect the physical existence of the postulated development clock of switching and we will attempt to map its chromosomal location. Using fetal or adult hybrids we will investigate the correlation between beta locus replication and the expression of fetal or adult globin genes. Second, we will investigate the control of the beta locus using hybrids containing human beta locus developmental mutants. These studies will include analyses of the functional effects of deletions of the Locus Activation Region; chromatin studies of deletion HPFH and delta beta thalassemia mutants; investigations of genetic heterogeneity and chromosomal mapping of non-deletion HPFH mutants. The proposed studies cannot be done with other experimental systems and are based on the fact that hybrids containing mutant loci can serve as surrogate cells of human erythroid cells. Third, we will use the hybrids as a model system for studying the control of the beta globin locus using homologous recombination. Questions such as the relationship between physical order of genes and order of developmental expression will be asked. Fourth, we will use subtraction hybridization of cDNA libraries from hybrids expressing the fetal or the adult globin program in order to clone genes coding for trans acting factors involved in globin gene switching. Fifth, we will continue our research on human erythroleukemia lines and we will use these lines as well as hybrids in order to study the developmental control of other than globin erythroid characters. Successful completion of the proposed studies will provide several insights on the control of globin genes during development and differentiation.

该赠款提案的长期目标是划定控制 在人类中从胎儿变为成年球蛋白形成的转换 发展。 为此，我们将应用体细胞杂交 我们开发的方法使我们能够对 正常的正常，突变或工程球蛋白基因的表达 染色体位置。 使用此系统，我们将首先研究 伽马在个体发育过程中切换的机理。 我们将尝试 获得明确的证据，表明在 跨我们将进行旨在检测物理存在的研究 开关的开发时钟，我们将尝试映射 其染色体位置。 使用胎儿或成人杂种，我们将调查 Beta基因座复制与胎儿表达之间的相关性 或成人球蛋白基因。 其次，我们将调查Beta的控制 使用含有人β基因座发展突变体的杂种基因座。 这些研究将包括对缺失功能效应的分析 基因座激活区域；缺失HPFH和 Delta beta thalassya突变体；研究遗传异质性和 非缺失HPFH突变体的染色体图。 提出的研究 无法使用其他实验系统来完成，并且是基于事实 含有突变基因座的杂种可以用作人类的替代细胞 红细胞细胞。 第三，我们将使用混合动力车作为模型系统 使用同源研究研究β球蛋白基因座的控制 重组。 诸如身体秩序之间的关系之类的问题 将询问基因和发育表达的顺序。 第四，我们 将使用杂种的cDNA库的减法杂交 表达胎儿或成人球蛋白程序以克隆基因 编码涉及球蛋白基因转换的反式作用因子。 第五， 我们将继续对人类红血球血压的研究，我们将 使用这些线和杂种来研究发展 除了球蛋白红细胞字符以外的其他控制。 成功完成 在拟议的研究中，将提供有关控制的几个见解 发育和分化过程中的球蛋白基因。