Jiang S W, Eberhardt N L
Department of Medicine, Mayo Clinic, Rochester, Minnesota 55905, USA.
J Biol Chem. 1995 Jun 9;270(23):13906-15. doi: 10.1074/jbc.270.23.13906.
Previous studies suggested that transcription enhancer factor-1 (TEF-1) was involved in mediating the human chorionic somatomammotropin (hCS) gene enhancer (CSEn) function (Jiang, S.-W., and Eberhardt, N. L. (1994) J. Biol. Chem. 269, 10384-10392). We now show that an unrelated protein (CSEF-1) found in BeWo and COS-1 cells binds to the GT-IIC enhanson in CSEn and is correlated with CSEn activity in these cells. TEF-1 and CSEF-1 were distinguished by differential migration as GT-IIC complexes, thermal stability, molecular mass, and cross-reactivity with chicken TEF-1 antibodies. TEF-1 and CSEF-1 bound to the GT-IIC and Sph-I/Sph-II enhansons with identical binding properties, and in vitro generated TEF-1 competed with CSEF-1 binding to the GT-IIC motif, suggesting that their actions might be mutually exclusive. Up- and down-regulation of TEF-1 levels by expression systems and antisense oligonucleotides demonstrated that TEF-1 inhibited the hCS promoter in a manner independent of the enhancer or a known TEF-1 DNA binding site. The data suggest that TEF-1 may provide a counter-regulatory stimulus to the actions of CSEF-1, which may be involved in mediating enhancer stimulatory activity.
先前的研究表明,转录增强因子-1(TEF-1)参与介导人绒毛膜生长催乳素(hCS)基因增强子(CSEn)的功能(蒋,S.-W.,和埃伯哈特,N. L.(1994年)《生物化学杂志》269,10384 - 10392)。我们现在发现,在BeWo和COS-1细胞中发现的一种不相关蛋白(CSEF-1)与CSEn中的GT-IIC增强子结合元件结合,并且与这些细胞中的CSEn活性相关。TEF-1和CSEF-1通过作为GT-IIC复合物的差异迁移、热稳定性、分子量以及与鸡TEF-1抗体的交叉反应性来区分。TEF-1和CSEF-1以相同的结合特性与GT-IIC和Sph-I/Sph-II增强子结合元件结合,并且体外产生的TEF-1与CSEF-1竞争结合GT-IIC基序,这表明它们的作用可能相互排斥。通过表达系统和反义寡核苷酸对TEF-1水平的上调和下调表明,TEF-1以独立于增强子或已知TEF-1 DNA结合位点的方式抑制hCS启动子。数据表明,TEF-1可能对CSEF-1的作用提供一种反调节刺激,而CSEF-1可能参与介导增强子刺激活性。