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首页> 外文期刊>Genomics >Molecular cloning of the mouse AMY-1 gene and identification of the synergistic activation of the AMY-1 promoter by GATA-1 and Sp1( small star, filled ).
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Molecular cloning of the mouse AMY-1 gene and identification of the synergistic activation of the AMY-1 promoter by GATA-1 and Sp1( small star, filled ).

机译:小鼠AMY-1基因的分子克隆以及由GATA-1和Sp1(小星,实心)识别的AMY-1启动子的协同激活。

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摘要

We have reported that a novel c-Myc binding protein, AMY-1, stimulated the transcription activity of c-Myc and was translocated from the cytoplasm to the nucleus in a c-Myc-dependent manner. AMY-1 works as an inducer of human K562 cell differentiation upon induction of AraC. To characterize the expression or functional importance of AMY-1, the genomic DNA of mouse AMY-1 was cloned and characterized. Both mouse and human genomic DNAs, the latter of which was retrieved from a human DNA database, comprise five exons spanning about 11 kb. To characterize the promoter of the mouse AMY-1 gene, a series of deletion constructs of the region upstream of the first ATG was linked to the luciferase gene, and their luciferase activities were measured in human HeLa and K562 cells. The results showed that Sp1 was essential for AMY-1 expression in both cell lines and that GATA-1 is also necessary in K562 cells. Sp1 in both cell lines and GATA-1 only in K562 cells were identified as proteins binding to these sites by amobility shift assay. Furthermore, it was found that GATA-1 stimulated AMY-1 expression synergistically with Sp1 in ectopically expressed insect cells and that both proteins were associated in K562 cells.
机译:我们已经报道了一种新型的c-Myc结合蛋白AMY-1,刺激了c-Myc的转录活性,并以c-Myc依赖的方式从细胞质转移到细胞核。 AMY-1在诱导AraC时可作为人类K562细胞分化的诱导剂。为了表征AMY-1的表达或功能重要性,克隆并表征了小鼠AMY-1的基因组DNA。小鼠和人类基因组DNA均包含五个外显子,跨度约为11 kb,后者是从人类DNA数据库中检索到的。为了表征小鼠AMY-1基因的启动子,将第一个ATG上游区域的一系列缺失构建体与荧光素酶基因连接,并在人HeLa和K562细胞中测量了它们的荧光素酶活性。结果表明,Sp1对于两种细胞系中的AMY-1表达都是必不可少的,而GATA-1在K562细胞中也是必需的。通过迁移率迁移分析,细胞系中的Sp1和仅K562细胞中的GATA-1被识别为与这些位点结合的蛋白质。此外,发现在异位表达的昆虫细胞中,GATA-1与Sp1协同刺激了AMY-1表达,并且两种蛋白质在K562细胞中都相关。

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