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一种稳定的转录复合物通过RNA聚合酶I指导小鼠核糖体RNA的合成。

A stable transcription complex directs mouse ribosomal RNA synthesis by RNA polymerase I.

作者信息

Cizewski V, Sollner-Webb B

出版信息

Nucleic Acids Res. 1983 Oct 25;11(20):7043-56. doi: 10.1093/nar/11.20.7043.

Abstract

Ribosomal RNA is synthesized from template molecules that are activated by a stable association with essential transcription factors. This activated template assembles prior to the onset of transcription as a preinitiation complex and factors remain firmly attached during active elongation as well. Sequential addition of differently marked rRNA genes to an S-100 mouse cell extract shows that the DNA binding factors of the stable complex are present in limiting quantities. They associate rapidly with template molecules and the resultant transcription complex remains intact over prolonged periods of incubation in the presence of competitor DNA. The resistance of the stable complex to the usual inhibitory effect of high DNA concentration suggests that more than one DNA binding factor recognizes the rDNA promoter region and is needed to direct faithful transcription. Finally, although the stable complex is specific for the rRNA initiation region, added vector sequences can neutralize nonspecific DNA binding components that are also present in the cell extract. This lowers the requirement for rDNA template and demonstrates that each activated rRNA gene can direct at least 10 rounds of elongation and reinitiation.

摘要

核糖体RNA是由与必需转录因子稳定结合而被激活的模板分子合成的。这种被激活的模板在转录开始前组装成预起始复合物,并且在活跃延伸过程中这些因子也会牢固附着。将不同标记的rRNA基因顺序添加到S - 100小鼠细胞提取物中表明,稳定复合物的DNA结合因子数量有限。它们与模板分子迅速结合,并且在存在竞争DNA的情况下长时间孵育时,所得转录复合物仍保持完整。稳定复合物对高DNA浓度通常的抑制作用具有抗性,这表明不止一种DNA结合因子识别rDNA启动子区域,并且需要它们来指导准确转录。最后,尽管稳定复合物对rRNA起始区域具有特异性,但添加的载体序列可以中和细胞提取物中也存在的非特异性DNA结合成分。这降低了对rDNA模板的需求,并表明每个被激活的rRNA基因可以指导至少10轮延伸和重新起始。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e019/326437/23c91b615d39/nar00365-0131-a.jpg

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