TRANS-ACTING FACTOR(S) CONTROLLING GLOBIN GENE EXPRESSION IN K562 CELLS

K562 细胞中控制珠蛋白基因表达的反式作用因子

• 批准号: 3940475
• 负责人: P GUTMAN
• 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/3940475
• 关键词: autoradiography; azacitidine; developmental genetics; gel electrophoresis; gel filtration chromatography; gene expression; gene interaction; genes; genetic manipulation; genetic regulation; genetic transcription; human tissue; ion exchange chromatography; messenger RNA; neoplastic cell culture for noncancer research; nucleic acid sequence; plasmids; radiotracer

K562 is an erythroleukemic cell line widely used as a model for the study of the control of human globin gene expression. These cells do not support transcription of the beta-globin gene (human adult pattern of expression) but do express transcripts of epsilon- and gamma-globin genes (human embryonic and fetal pattern) at very high levels when exposed to a number of inducing agents. Results from this and other laboratories suggest that the control of this pattern of expression is mediated by the presence and/or absence of trans-acting factors which exert their action on sequences corresponding to the promoters of these genes. Sequence specific DNA binding proteins acting on cis-regulatory control elements have been hypothersized to be key elements in eukaryotic gene transcription, and even though considerable progress has been made in their isolation, DNA binding proteins with affinity for the human globin gene promoters have not yet been identified. We have chosen to study the interaction of these factors with DNA sequences belonging to the epsilon-gene promoter. The methodology used DNA sequences belonging to the epsilon-gene promoter. The methodology used included DNase footprinting and the gel retardation assay. By the former, two protective patterns have been detected surrounding the -500 and - 260 nucleotide regions, and by the latter multiple DNA binding activities have been shown, some of which possess specificity. Experiments leading to the detection of the exact sites to which these elements bind are now under course using a combination of these two methods. Further studies will be undertaken to show the functional significance of these factors. Characterization of such factors is crucial in understanding the mechanisms underlying the transcriptional control of human globin genes.

K562 是一种广泛用作模型的红白血病细胞系 人类珠蛋白基因表达控制的研究。 这些 细胞不支持β-珠蛋白基因（人类 成人表达模式），但确实表达 epsilon- 的转录本 和伽马珠蛋白基因（人类胚胎和胎儿模式） 当暴露于多种诱导剂时，其水平非常高。 该实验室和其他实验室的结果表明，控制 这种表达模式的存在是由存在和/或 缺乏发挥其作用的反式作用因子 对应于这些基因的启动子的序列。 作用于顺式调控的序列特异性 DNA 结合蛋白 控制元件被假设为关键元件 真核基因转录，尽管相当大 DNA结合蛋白的分离已取得进展 与人类珠蛋白基因启动子具有亲和力尚未 已被识别。 我们选择研究这些之间的相互作用 具有属于 epsilon 基因的 DNA 序列的因子 发起人。 该方法使用的 DNA 序列属于 ε-基因启动子。 使用的方法包括 DNase 足迹法和凝胶阻滞测定。 就前者而言，两个 已在 -500 和 - 周围检测到保护图案 260个核苷酸区域，并由后者与多个DNA结合 已经显示了一些活动，其中一些活动具有特殊性。 导致检测到的确切位点的实验 这些元素绑定现在正在使用以下组合 这两种方法。 将进行进一步的研究以表明 这些因素的功能意义。 表征 这些因素对于理解机制至关重要 人类珠蛋白基因转录控制的基础。