The transcriptional co-activator TAZ interacts differentially with transcriptional enhancer factor-1 (TEF-1) family members
The transcriptional co-activator TAZ interacts differentially with transcriptional enhancer factor-1 (TEF-1) family members
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DOI：
10.1042/bj20041434
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发表时间：
2005-05-15
影响因子：
4.1
通讯作者：
Farrance, KG
中科院分区：
文献类型：
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作者：
Mahoney, WM;Hong, JH;Farrance, KG
高度相关的TEF-1（转录增强因子-1）家族成员（也称为TEAD， TEF-1、TEC1、ABAA结构域）结合MCAT（肌肉C、A和T位点）和在心脏、骨骼肌、平滑肌、胎盘和神经嵴中活跃的启动子中富含A[f]的位点。TEF-1活性受转录辅助因子[p160， TONDU (Vg1-1, Vestigial-like protein-1)， Vg1-2和YAP65 (Yes-associated protein 6- 5kda)]的相互作用调节。强转录共激活因子YAP65与所有TEF-1家族成员相互作用，并且由于YAP65与TAZ（带pdz结合基序的转录共激活因子）相关，我们想确定TAZ是否也与TEF-1家族成员相互作用。在本研究中，我们通过GST（谷胱甘肽s -转移酶）下拉实验、共免疫沉淀和改良的哺乳动物双杂交实验表明，TEF-1在体外和体内与TAZ相互作用。用纯化的TEF-1和GST-TAZ融合蛋白进行的电泳迁移试验表明，TAZ与结合在MCAT DNA上的TEF-1相互作用。TAZ可以与内源性TEF-1蛋白相互作用，因为外源性TAZ激活了mcat依赖的报告启动子。像YAP65一样，TAZ与所有四个TEF-1家族成员相互作用。随着[S-35]TEF-1和[S-35]RTEF-1（相关TEF-1）含量的增加，GST下拉试验表明，TAZ与TEF-1的相互作用比与RTEF-1的相互作用更有效。这种差异的相互作用也扩展到TEF-1和RTEF-1与TAZ在体内的相互作用，通过改进的哺乳动物双杂交实验进行了分析。这些数据表明，TEF-1蛋白与转录共激活因子的差异关联可能调节TEF-1家族成员的活性。
Members of the highly related TEF-1 (transcriptional enhancer factor-1) family (also known as TEAD, for TEF-1, TEC1, ABAA domain) bind to MCAT (muscle C, A and T sites) and A[F-rich sites in promoters active in cardiac, skeletal and smooth muscle, placenta, and neural crest. TEF-1 activity is regulated by interactions with transcriptional co-factors [p160, TONDU (Vg1-1, Vestigial-like protein-1), Vg1-2 and YAP65 (Yes-associated protein 6-5 kDa)]. The strong transcriptional co-activator YAP65 interacts with all TEF-1 family members, and, since YAP65 is related to TAZ (transcriptional co-activator with PDZ-binding motif), we wanted to determine if TAZ also interacts with members of the TEF-1 family. In the present study, we show by GST (glutathione S-transferase) pull-down assays, by co-immunoprecipitation and by modified mammalian two-hybrid assays that TEF-1 interacts with TAZ in vitro and in vivo. Electrophoretic mobility-shift assays with purified TEF-1 and GST-TAZ fusion protein showed that TAZ interacts with TEF-1 bound to MCAT DNA. TAZ can interact with endogenous TEF-1 proteins, since exogenous TAZ activated MCAT-dependent reporter promoters. Like YAP65, TAZ interacted with all four TEF-1 family members. GST pull-down assays with increasing amounts of [S-35]TEF-1 and [S-35]RTEF-1 (related TEF-1) showed that TAZ interacts more efficiently with TEF-1 than with RTEF-1. This differential interaction also extended to the interaction of TEF-1 and RTEF-1 with TAZ in vivo, as assayed by a modified mammalian two-hybrid experiment. These data show that differential association of TEF-1 proteins with transcriptional co-activators may regulate the activity of TEF-1 family members.