Duyndam M C, van Dam H, van der Eb A J, Zantema A
Laboratory for Molecular Carcinogenesis, Sylvius Laboratory, Leiden University, The Netherlands.
J Virol. 1996 Sep;70(9):5852-9. doi: 10.1128/JVI.70.9.5852-5859.1996.
The adenovirus 12S E1A protein can stimulate the activity of the c-jun promoter through a conserved region 1 (CR1)-dependent mechanism. The effect is mediated by two AP-1/ATF-like elements, jun1 and jun2, that preferentially bind c-Jun-ATF-2 heterodimers. In this study, we show that the ATF-2 component of the c-Jun-ATF-2 heterodimer is the primary target for 12S E1A: 12S E1A can enhance the transactivating activity of the N terminus of ATF-2 when fused to a heterologous DNA-binding domain, whereas the transactivating activity of the c-Jun N terminus is not significantly affected. Activation of the ATF-2 N terminus by 12S E1A is dependent on CR1. In the context of the 13S E1A protein, CR1 and CR3 can both contribute to activation of ATF-2, and their relative contributions are dependent on the cell type. In contrast to activation of ATF-2 by stress-inducing agents, CR1-dependent activation of ATF-2 was found not to depend strictly on the presence of threonines 69 and 71 in the N terminus of ATF-2, which are targets for phosphorylation by stress-activated protein kinases (SAPKs). In agreement with this observation, we did not observe phosphorylation of threonines 69 and 71 or constitutively enhanced SAPK activity in E1A- plus E1B-transformed cell lines. These data suggest that CR1-dependent activation of ATF-2 by 12S E1A does not require phosphorylation of threonines 69 and 71 by SAPK.
腺病毒12S E1A蛋白可通过依赖保守区域1(CR1)的机制刺激c-jun启动子的活性。这种效应由两个AP-1/ATF样元件jun1和jun2介导,它们优先结合c-Jun-ATF-2异二聚体。在本研究中,我们表明c-Jun-ATF-2异二聚体的ATF-2组分是12S E1A的主要作用靶点:当与异源DNA结合结构域融合时,12S E1A可增强ATF-2 N端的反式激活活性,而c-Jun N端的反式激活活性未受到显著影响。12S E1A对ATF-2 N端的激活依赖于CR1。在13S E1A蛋白的情况下,CR1和CR3均可促进ATF-2的激活,它们的相对贡献取决于细胞类型。与应激诱导剂对ATF-2的激活不同,发现CR1依赖的ATF-2激活并不严格依赖于ATF-2 N端苏氨酸69和71的存在,而这两个位点是应激激活蛋白激酶(SAPK)磷酸化的靶点。与这一观察结果一致,我们在E1A和E1B共转化的细胞系中未观察到苏氨酸69和71磷酸化或持续增强的SAPK活性。这些数据表明,12S E1A通过CR1依赖机制激活ATF-2不需要SAPK对苏氨酸69和71进行磷酸化。