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首页> 外文期刊>Molecular and Cellular Biology >Linker scanning mutagenesis of the 5'-flanking region of the mouse beta-major-globin gene: sequence requirements for transcription in erythroid and nonerythroid cells.
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Linker scanning mutagenesis of the 5'-flanking region of the mouse beta-major-globin gene: sequence requirements for transcription in erythroid and nonerythroid cells.

机译：小鼠β-主要球蛋白基因5'侧翼区域的接头扫描诱变：在红系和非红系细胞中转录的序列要求。

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摘要

We analyzed the sequences required for transcription of the mouse beta-major-globin gene by introducing deletion and linker scanning mutations into the 5'-flanking region and then studying the effects of these mutations on beta-globin gene transcription in a HeLa cell transient expression assay or after stable introduction into mouse erythroleukemia cells. Consistent with earlier studies, we found that three distinct regions upstream from the RNA capping site are required for efficient beta-globin gene transcription in HeLa cells: the ATA box located 30 base pairs upstream from the mRNA capping site (-30), the CCAAT box located at -75, and the distal sequence element CCACACCC located at -90. In the ATA and CAAT box regions, the sequences necessary for efficient transcription extend beyond the limits of the canonical sequences. Mutations in the sequences located between the three transcriptional control elements do not significantly affect transcription in HeLa cells. Although the promoter defined in HeLa cell transfection experiments is also required for efficient transcription in mouse erythroleukemia cells, none of the mutations tested affects the regulation of beta-globin gene transcription during mouse erythroleukemia cell differentiation. Thus, DNA sequences downstream from the mRNA cap site appear to be sufficient for the regulation of beta-globin gene expression during the differentiation of mouse erythroleukemia cells in culture.
机译：我们通过将缺失和接头扫描突变引入5'侧翼区域，然后研究这些突变对HeLa细胞瞬时表达中β-球蛋白基因转录的影响，分析了小鼠β-主要球蛋白基因转录所需的序列检测或稳定引入小鼠红白血病细胞后。与早期研究一致，我们发现，在HeLa细胞中有效的β-珠蛋白基因转录需要RNA帽位点上游三个不同区域：ATA盒位于mRNA帽位点（-30）上游30个碱基对，CCAAT框位于-75，远端序列元素CCACACCC位于-90。在ATA和CAAT框区域中，有效转录所必需的序列超出了规范序列的范围。位于三个转录控制元件之间的序列中的突变不会显着影响HeLa细胞中的转录。尽管在小鼠红白血病细胞中进行有效转录也需要在HeLa细胞转染实验中定义的启动子，但测试的任何突变均不会影响小鼠红白血病细胞分化过程中β-珠蛋白基因转录的调控。因此，在培养的小鼠红白血病细胞分化过程中，mRNA帽位点下游的DNA序列似乎足以调节β-珠蛋白基因的表达。

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