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首页> 外文期刊>Molecular and Cellular Biology >Recruitment of an RNA Polymerase II Complex Is Mediated by the Constitutive Activation Domain in CREB, Independently of CREB Phosphorylation
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Recruitment of an RNA Polymerase II Complex Is Mediated by the Constitutive Activation Domain in CREB, Independently of CREB Phosphorylation

机译：RNA聚合酶II复合物的招聘由CREB中的本构激活域介导，与CREB磷酸化无关

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摘要

The cAMP response element binding protein (CREB) is a bifunctional transcription activator, exerting its effects through a constitutive activation domain (CAD) and a distinct kinase inducible domain (KID), which requires phosphorylation of Ser-133 for activity. Both CAD and phospho-KID have been proposed to recruit polymerase complexes, but this has not been directly tested. Here, we show that the entire CREB activation domain or the CAD enhanced recruitment of a complex containing TFIID, TFIIB, and RNA polymerase II to a linked promoter. The nuclear extracts used mediated protein kinase A (PKA)-inducible transcription, but phosphorylation of CRG (both of the CREB activation domains fused to the Gal4 DNA binding domain) or KID-G4 did not mediate recruitment of a complex, and mutation of the PKA site in CRG abolished transcription induction by PKA but had no effect upon recruitment. The CREB-binding protein (CBP) was not detected in the recruited complex. Our results support a model for transcription activation in which the interaction between the CREB CAD and hTAFII130 of TFIID promotes the recruitment of a polymerase complex to the promoter.
机译：cAMP反应元件结合蛋白（CREB）是一种双功能转录激活剂，通过组成型激活域（CAD）和独特的激酶诱导域（KID）发挥其作用，后者需要将Ser-133磷酸化才能发挥活性。已经提出了CAD和磷酸化KID来募集聚合酶复合物，但是尚未对此进行直接测试。在这里，我们显示了整个CREB激活域或CAD增强了包含TFIID，TFIIB和RNA聚合酶II的复合物向连接的启动子的募集。核提取物使用介导的蛋白激酶A（PKA）诱导的转录，但CRG（两个CREB激活结构域与Gal4 DNA结合结构域融合）或KID-G4的磷酸化均不介导复合物的募集，并且该蛋白的突变CRG中的PKA位点废除了PKA的转录诱导作用，但对募集没有影响。在募集的复合物中未检测到CREB结合蛋白（CBP）。我们的结果支持转录激活模型，其中CREB ​​CAD和TFIID的hTAFII130之间的相互作用促进了聚合酶复合物向启动子的募集。

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