CHROMATIN STRUCTURE IN REGULATION OF MAMMALIAN GENE EXPRESSION

哺乳动物基因表达调节中的染色质结构

• 批准号: 5201910
• 负责人: A DEAN
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/5201910
• 关键词: chromatin; developmental genetics; gene expression; genetic enhancer element; genetic promoter element; genetic regulation; genetic transcription; globin; human genetic material tag; mammalian embryology; nucleic acid structure; nucleosomes; tissue /cell culture

The embryonic epsilon-globin gene is the first member of the beta-globin family of genes to be expressed during development. Expression of the beta-like-globin genes is under the control of the beta-globin locus control region (LCR), encompassing erythroid specific DNase I hypersensitive sites (HS) 5-18 kilobases upstream of the epsilon-globin gene. The LCR has long range effects on the chromatin structure of the beta-globin locus, and it activates transcription of the different globin gene promoters as the genes are expressed sequentially during development. Much of our current understanding of the process of gene activation has been gained from in vitro studies where DNA is not assembled with histones and other chromosomal proteins as it is in the nucleus. To study the activation of a gene in its natural context of chromatin in vivo we have used minichromosomes containing the human epsilon-globin gene and portions of the beta globin LCR enhancer. Expression of hue minichromosomal epsilon-globin gene in K562 human erythroid cells is completely dependent on the presense if the beta- globin LCR in cis. Using restriction endonuclease access, and indirect end labelling experiments, we have observed that the promoter and 5' proximal structural regions of the minichromosomal epsilon-globin gene are covered by an array of positioned nucleosomes. Both types of studies suggest alteration or loss of a single promoter-proximal nucleosome upon activation of the gene for transcription. Regulated expression of the epsilon-globin gene on the minichromsomes offers a means too study at high resolution, the promoter-enhancer interaction of an activated gene assembled in vivo into chromatin.

胚胎表型珠蛋白基因是β-珠蛋白的第一个成员。 在发育过程中要表达的基因家族。的表达 类β珠蛋白基因受β-珠蛋白基因控制 控制区(LCR)，包括红系特异性DNA酶I 表珠蛋白上游5-18kb的超敏位点 吉恩。LCR对细胞的染色质结构有长程影响 β-珠蛋白基因座，它激活不同珠蛋白的转录 基因启动子作为基因在过程中顺序表达 发展。我们目前对基因过程的大部分理解 激活是从体外研究中获得的，而DNA不是 与组蛋白和其他染色体蛋白质组装在一起 原子核。研究基因在其自然环境中的激活 染色质在体内我们已经使用了含有人的微小染色体 Epsilon-珠蛋白基因和部分β珠蛋白LCR增强子。 HUE微小染色体表型珠蛋白基因在K562人中的表达 红系细胞完全依赖于存在，如果β- 顺式中的珠蛋白LCR。使用限制性内切酶访问，和间接 结束标记实验，我们观察到启动子和5‘ 微染色体表型珠蛋白基因的近端结构区 被一系列定位的核小体所覆盖。这两种类型的研究 提示单个启动子-近端核小体的改变或丢失 激活基因以进行转录。受调控的基因表达 微小染色体上的Epsilon-珠蛋白基因也提供了一种研究手段 高分辨率，激活基因的启动子-增强子相互作用 在体内组装成染色质。