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与β珠蛋白主基因启动子相互作用的核蛋白在诱导后12小时内开始在MEL细胞中积累,并且该启动子的RNA拷贝在体外能成功竞争它们的结合。

Nuclear proteins that interact with the beta maj globin promoter start to accumulate in MEL cells within 12 hours of induction and RNA copies of the promoter successfully compete their binding in vitro.

作者信息

Sun-Hoffman L, Winicov I

机构信息

Department of Microbiology, University of Nevada, Reno, USA.

出版信息

Mol Cell Biochem. 1995 Apr 26;145(2):159-68. doi: 10.1007/BF00935488.

Abstract

The induction of differentiation in mouse erythroleukemia (MEL) cells by dimethylsulfoxide (DMSO) is characterized by increased transcription of globin genes. We have determined that DMSO treated cells increase the levels of nuclear factors capable of overall interactions with the beta maj globin promoter during the initial 24 h post induction, as measured by gel mobility analysis. Two unprocessed beta maj globin mRNA precursors, which are present in MEL cell nuclei early in differentiation, were previously shown to contain the 5' promoter flanking region, and thereby provided the nucleus with a pool of regulatory sequences in multiple RNA copies. We have studied the effect of RNA copies of the promoter region on binding interactions between DNA sequences of the beta maj globin promoter and nuclear factors that interact with these sequences. The promoter region RNA transcripts competed effectively for DNA binding proteins in vitro, while the antisense RNA from the same region did not. The most pronounced competition was observed with proteins from 12 h after DMSO induction, when the concentration of the DNA binding proteins was still increasing. Since the 'upstream' transcripts predominate at 12 h after DMSO induction, these results indicate that the promoter region transcripts may influence the equilibrium of binding between the beta maj globin promoter and the nuclear factors that bind to this region during DMSO induction.

摘要

二甲基亚砜(DMSO)诱导小鼠红白血病(MEL)细胞分化的特征是珠蛋白基因转录增加。我们已经确定,通过凝胶迁移分析测量,在诱导后的最初24小时内,经DMSO处理的细胞增加了能够与βmaj珠蛋白启动子进行全面相互作用的核因子水平。先前已证明,在分化早期存在于MEL细胞核中的两种未加工的βmaj珠蛋白mRNA前体包含5'启动子侧翼区域,从而为细胞核提供了多个RNA拷贝形式的调控序列库。我们研究了启动子区域的RNA拷贝对βmaj珠蛋白启动子的DNA序列与与这些序列相互作用的核因子之间结合相互作用的影响。启动子区域的RNA转录本在体外有效地竞争DNA结合蛋白,而来自同一区域的反义RNA则没有。在DMSO诱导后12小时观察到与蛋白质的最明显竞争,此时DNA结合蛋白的浓度仍在增加。由于“上游”转录本在DMSO诱导后12小时占主导地位,这些结果表明启动子区域转录本可能会影响DMSO诱导期间βmaj珠蛋白启动子与结合到该区域的核因子之间结合的平衡。

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