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牛α-骨骼肌肌动蛋白基因的核苷酸序列、结构及转录调控的初步研究

The nucleotide sequence, structure, and preliminary studies on the transcriptional regulation of the bovine alpha skeletal actin gene.

作者信息

Davey H W, Kelly J K, Wildeman A G

机构信息

Department of Molecular Biology and Genetics, University of Guelph, Ontario, Canada.

出版信息

DNA Cell Biol. 1995 Jul;14(7):609-18. doi: 10.1089/dna.1995.14.609.

Abstract

The promoters of mammalian striated muscle actin gene contain binding sites for a number of transcription factors. Examples are the CArG boxes, which bind a protein identical to or related to serum response factor (SRF), E boxes, which bind myogenic determination factors such as MyoD and myogenin, and -CCGCCC- motifs, which bind the transcription factor Sp1. To date, the only mammalian sequences isolated and analyzed are from rodent and human. We have now isolated and sequenced the bovine gene encoding alpha skeletal actin, including almost 3 kb of 5'-flanking region. When compared to the human and rodent genes (the only ones previously cloned and for which 5'-flanking sequences to only approximately -750 are known), there was the expected conservation in the coding region. A comparison of the promoter regions indicated that the bovine gene has three CArG boxes in the 5'-flanking region in positions identical to those in other species. The bovine proximal promoter is unique from those of human and rodent in that it has only one E box in the vicinity of the TATA box, near -350, whereas the other mammals have three. Far upstream sequences reveal clusters of E boxes near -2,500 and -1,500. A minimal promoter element, to -297, which has no E boxes, is sufficient to activate transcription in myotubes derived from rat L6 and mouse C2C12 myoblasts.

摘要

哺乳动物横纹肌肌动蛋白基因的启动子含有多个转录因子的结合位点。例如,CArG框可结合与血清反应因子(SRF)相同或相关的蛋白质;E框可结合生肌决定因子,如MyoD和肌细胞生成素;-CCGCCC-基序可结合转录因子Sp1。迄今为止,分离和分析的仅有的哺乳动物序列来自啮齿动物和人类。我们现已分离并测序了编码α骨骼肌肌动蛋白的牛基因,包括近3 kb的5'侧翼区。与人类和啮齿动物基因(此前仅有的已克隆基因,其5'侧翼序列仅已知约-750)相比,编码区存在预期的保守性。对启动子区域的比较表明,牛基因在5'侧翼区有三个CArG框,其位置与其他物种相同。牛近端启动子与人类和啮齿动物的不同,在于它在TATA框附近(约-350)只有一个E框,而其他哺乳动物有三个。远上游序列在-2500和-1500附近显示出E框簇。一个至-297的最小启动子元件,其没有E框,足以激活源自大鼠L6和小鼠C2C12成肌细胞的肌管中的转录。

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