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克隆的RNA聚合酶II转录因子IID在体外选择RNA聚合酶III来转录人类U6基因。

The cloned RNA polymerase II transcription factor IID selects RNA polymerase III to transcribe the human U6 gene in vitro.

作者信息

Lobo S M, Lister J, Sullivan M L, Hernandez N

机构信息

Cold Spring Harbor Laboratory, New York 11724.

出版信息

Genes Dev. 1991 Aug;5(8):1477-89. doi: 10.1101/gad.5.8.1477.

Abstract

Although the human U2 and U6 snRNA genes are transcribed by different RNA polymerases (i.e., RNA polymerases II and III, respectively), their promoters are very similar in structure. Both contain a proximal sequence element (PSE) and an octamer motif-containing enhancer, and these elements are interchangeable between the two promoters. The RNA polymerase III specificity of the U6 promoter is conferred by a single A/T-rich element located around position -25. Mutation of the A/T-rich region converts the U6 promoter into an RNA polymerase II promoter, whereas insertion of the A/T-rich region into the U2 promoter converts that promoter into an RNA polymerase III promoter. We show that this A/T-rich element can be replaced by a number of TATA boxes derived from mRNA promoters transcribed by RNA polymerase II with little effect on RNA polymerase III transcription. Furthermore, the cloned RNA polymerase II transcription factor TFIID both binds to the U6 A/T-rich region and directs accurate RNA polymerase III transcription in vitro. Mutations in the U6 A/T-rich region that convert the U6 promoter into an RNA polymerase II promoter also abolish TFIID binding. Together, these observations suggest that in the human snRNA promoters, unlike in mRNA promoters, binding of TFIID directs the assembly of RNA polymerase III transcription complexes, whereas the lack of TFIID binding results in the assembly of RNA polymerase II snRNA transcription complexes.

摘要

尽管人类U2和U6小核RNA基因分别由不同的RNA聚合酶转录(即分别由RNA聚合酶II和III转录),但其启动子在结构上非常相似。两者都包含一个近端序列元件(PSE)和一个含八聚体基序的增强子,并且这些元件在两个启动子之间是可互换的。U6启动子的RNA聚合酶III特异性由位于约-25位置的单个富含A/T的元件赋予。富含A/T区域的突变将U6启动子转变为RNA聚合酶II启动子,而将富含A/T的区域插入U2启动子则将该启动子转变为RNA聚合酶III启动子。我们发现,这个富含A/T的元件可以被许多源自由RNA聚合酶II转录的mRNA启动子的TATA框所取代,而对RNA聚合酶III转录影响很小。此外,克隆的RNA聚合酶II转录因子TFIID既能与U6富含A/T的区域结合,又能在体外指导精确的RNA聚合酶III转录。将U6启动子转变为RNA聚合酶II启动子的U6富含A/T区域的突变也会消除TFIID的结合。总之,这些观察结果表明,在人类小核RNA启动子中,与mRNA启动子不同,TFIID的结合指导RNA聚合酶III转录复合物的组装,而缺乏TFIID的结合则导致RNA聚合酶II小核RNA转录复合物的组装。

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