Choi O R, Engel J D
Nature. 1986;323(6090):731-4. doi: 10.1038/323731a0.
The chicken adult beta-globin gene is one of the more intensively investigated developmentally regulated loci in higher eukaryotes. Detailed molecular analysis of the locus allows precise examination of the chromosomal changes that occur on activation of the gene during erythroid maturation. The best studied of these changes are the acquisition of DNase I hypersensitivity, developmentally correlated alteration of CpG-specific cytosine methylation patterns and in vitro assembly of erythroid-specific protein complexes 5' to the gene that mimics in vivo creation of the 5' DNase I hypersensitive 'region' lying 60 to 260 nucleotides 5' to the beta-globin cap site in red blood cell chromatin. Here we demonstrate that proximal beta-globin DNA sequences lying greater than 112 base pairs (bp) 5' to the cap site are not involved in determining the erythroid-specific induction characteristics of this gene in transient expression assays, whereas an enhancer sequence within a 300-bp PvuII fragment lying approximately 400 nucleotides 3' to the polyadenylation signal is intimately involved in determining the erythroid cell specificity and correct time of induction of beta-globin transcription during red cell maturation.
鸡的成人β-珠蛋白基因是高等真核生物中研究较为深入的发育调控基因座之一。对该基因座进行详细的分子分析,可以精确检测在红细胞成熟过程中基因激活时发生的染色体变化。其中研究得最为透彻的变化包括获得对DNase I的超敏感性、与发育相关的CpG特异性胞嘧啶甲基化模式的改变,以及在基因5'端体外组装红细胞特异性蛋白复合物,这种组装模拟了在红细胞染色质中位于β-珠蛋白帽位点5'端60至260个核苷酸处的5' DNase I超敏感“区域”的体内形成过程。在此,我们证明在瞬时表达试验中,位于帽位点5'端大于112个碱基对(bp)的近端β-珠蛋白DNA序列不参与决定该基因的红细胞特异性诱导特性,而位于多聚腺苷酸化信号3'端约400个核苷酸处的一个300 bp PvuII片段内的增强子序列,在决定红细胞特异性以及红细胞成熟过程中β-珠蛋白转录正确的诱导时间方面密切相关。