Majello B, De Luca P, Lania L
Department of Genetics, Molecular and General Biology, University of Naples "Federico II," via Mezzocannone 8, 80134 Naples 10, Italy.
J Biol Chem. 1997 Feb 14;272(7):4021-6. doi: 10.1074/jbc.272.7.4021.
Sp3 is a member of the Sp family of transcription factors and binds to DNA with affinity and specificity comparable to that of Sp1. We demonstrate that Sp3 is a bifunctional transcription factor that can both activate and repress transcription. Gene fusion experiments in mammalian cells demonstrate that the Sp3 activation potential is distributed over an extensive glutamine-rich N-terminal region, whereas the repressor activity has been mapped in a 72-amino acid region located at the 5' of the zinc finger DNA-binding domain. We demonstrated that the repression activity is strictly dependent on the context of the DNA-binding sites bound by Sp3. We found that Sp3 represses transcription of promoters bearing multiple GAL4 DNA-binding sites, whereas it activates isogenic reporters containing a single GAL4-binding site. Transfection experiments in Drosophila cells that lack endogenous Sp activity demonstrated that Sp3 does not possess an active repression domain that can function in insect cells, rather it is a weak transcriptional activator of the c-myc promoter. Our results strongly suggest that Sp3 is a dual-function regulator whose activity is dependent upon both the promoter and the cellular context.
Sp3是转录因子Sp家族的成员,它与DNA结合的亲和力和特异性与Sp1相当。我们证明Sp3是一种双功能转录因子,既能激活转录,也能抑制转录。哺乳动物细胞中的基因融合实验表明,Sp3的激活潜能分布在一个广泛的富含谷氨酰胺的N端区域,而其抑制活性则定位在位于锌指DNA结合结构域5'端的一个72个氨基酸的区域。我们证明抑制活性严格依赖于Sp3结合的DNA结合位点的背景。我们发现Sp3抑制带有多个GAL4 DNA结合位点的启动子的转录,而它激活含有单个GAL4结合位点的同基因报告基因。在缺乏内源性Sp活性的果蝇细胞中的转染实验表明,Sp3不具有能在昆虫细胞中起作用的活性抑制结构域,相反,它是c-myc启动子的弱转录激活剂。我们的结果强烈表明Sp3是一种双功能调节因子,其活性取决于启动子和细胞背景。