Institute for Human Genetics, Jena University Hospital, Germany.
FEBS J. 2013 Apr;280(8):1861-8. doi: 10.1111/febs.12211. Epub 2013 Mar 18.
The regulation of gene repression by corepressors is a controlled process. Surface-enhanced laser desorption ionization MS proteomic analysis and a yeast two-hybrid screen showed independently that the corepressor Alien interacts with the CREB-binding protein (CBP) coactivator. This interaction was further confirmed by coimmunoprecipitation and glutathione S-transferase pull-down experiments, suggesting that Alien interacts in vivo and in vitro with the histone acetyltransferase (HAT) coactivators CBP and its paralog p300. Acetylation detection experiments indicated that Alien is acetylated in vivo. Furthermore, Alien interacts with the central region of CBP/p300 containing the HAT domain and becomes acetylated in vitro. When an inhibitor of CBP/p300 HAT activity was employed, the Alien-mediated silencing was enhanced. Thus, these findings suggest crosstalk between corepressors and coactivators, and indicate fine-tuning of corepressor function by post-translational modification through corepressor acetylation.
p300 binds to Alien α by pull down (View interaction) Alien α physically interacts with CBP by two hybrid (View interaction) Alien α physically interacts with MLK2 by two hybrid (View interaction) p300 acetylates Alien α by acetylation assay (View interaction) Alien α physically interacts with NAP1 by two hybrid (View interaction) Alien α physically interacts with TAFI68 by two hybrid (View interaction) Alien α physically interacts with CBP by anti bait coimmunoprecipitation (View Interaction: 1, 2, 3) Alien α binds to CBP by pull down (View interaction).
核心抑制剂对基因抑制的调控是一个受控制的过程。表面增强激光解吸电离质谱蛋白质组分析和酵母双杂交筛选独立表明，核心抑制剂 Alien 与 CREB 结合蛋白 (CBP) 共激活剂相互作用。共免疫沉淀和谷胱甘肽 S-转移酶下拉实验进一步证实了这一相互作用，表明 Alien 在体内和体外均与组蛋白乙酰转移酶 (HAT) 共激活剂 CBP 及其旁系同源物 p300 相互作用。乙酰化检测实验表明，Alien 在体内被乙酰化。此外，Alien 与包含 HAT 结构域的 CBP/p300 的中心区域相互作用，并在体外被乙酰化。当使用 CBP/p300 HAT 活性抑制剂时，Alien 介导的沉默增强。因此，这些发现表明共抑制剂和共激活剂之间存在串扰，并表明通过核心抑制剂乙酰化等翻译后修饰来精细调节核心抑制剂的功能。
p300 通过下拉与 Alien α 结合（查看相互作用）Alien α 通过双杂交与 CBP 物理相互作用（查看相互作用）Alien α 通过双杂交与 MLK2 物理相互作用（查看相互作用）p300 通过乙酰化测定法乙酰化 Alien α（查看相互作用）Alien α 通过双杂交与 NAP1 物理相互作用（查看相互作用）Alien α 通过双杂交与 TAFI68 物理相互作用（查看相互作用）Alien α 通过抗诱饵共免疫沉淀与 CBP 物理相互作用（查看相互作用：1、2、3）Alien α 通过下拉与 CBP 结合（查看相互作用）。
