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K562细胞β-珠蛋白基因区域的核酸酶超敏反应。

Nuclease hypersensitivity in the beta-globin gene region of K562 cells.

作者信息

Cao S X, Schechter A N

机构信息

Laboratory of Chemical Biology, National Institute of Diabetes, and Digestive and Kidney Diseases, Bethesda, Maryland 20892.

出版信息

Eur J Biochem. 1988 May 2;173(3):517-22. doi: 10.1111/j.1432-1033.1988.tb14028.x.

Abstract

We have investigated chromatin structure in the beta-globin gene region of the K562 human erythroleukemic cell line by using S1 and DNase I nuclease sensitivity assays. Despite the lack of beta-globin gene expression in these cells, we find nuclease-hypersensitive sites to these enzymes in its 5' and 3' flanking regions in K562 chromatin. This result is in contrast to previous reports in which no hypersensitive sites were found in the immediate vicinity of this gene. In the 3' region, one major hypersensitive site at 0.9 kpb 3' and three minor hypersensitive sites at 0.7 kbp, 0.5 kbp 3' and 0.2 kbp 5' of the polyadenylation site were observed; these sites are very similar to those found in fetal liver and adult bone marrow cells in which the beta-globin gene is expressed. We find hypersensitive sites to both enzymes in the 5' region of the beta-globin gene: a major site 0.8 kbp 5' to the cap site, and two minor sites 1.2 and 1.5 kbp 5' to the cap site. The -0.8 kbp site is also present in plasmids containing the beta-globin gene. Our results suggest that the lack of beta-globin gene expression may be related to the lack of hypersensitivity sites in the immediate (150 bp) 5' flanking region of the beta-globin gene, as occurs in other active globin genes.

摘要

我们通过使用S1和DNase I核酸酶敏感性测定法,研究了K562人红白血病细胞系β-珠蛋白基因区域的染色质结构。尽管这些细胞中缺乏β-珠蛋白基因表达,但我们在K562染色质中该基因的5'和3'侧翼区域发现了对这些酶敏感的核酸酶超敏位点。这一结果与之前的报道相反,在之前的报道中该基因紧邻区域未发现超敏位点。在3'区域,观察到在多聚腺苷酸化位点下游0.9 kpb处有一个主要超敏位点,在其下游0.7 kbp、0.5 kbp和上游0.2 kbp处有三个次要超敏位点;这些位点与在表达β-珠蛋白基因的胎儿肝脏和成人骨髓细胞中发现的位点非常相似。我们在β-珠蛋白基因的5'区域发现了对这两种酶都敏感的超敏位点:在帽位点上游0.8 kbp处有一个主要位点,在帽位点上游1.2和1.5 kbp处有两个次要位点。-0.8 kbp位点也存在于含有β-珠蛋白基因的质粒中。我们的结果表明,β-珠蛋白基因表达的缺乏可能与β-珠蛋白基因紧邻(150 bp)的5'侧翼区域缺乏超敏位点有关,就像在其他活跃的珠蛋白基因中那样。

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