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由延伸因子TFIIS控制的RNA聚合酶II转录保真度

Fidelity of RNA polymerase II transcription controlled by elongation factor TFIIS.

作者信息

Jeon C, Agarwal K

机构信息

Department of Biochemistry and Molecular Biology, University of Chicago, IL 60637, USA.

出版信息

Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13677-82. doi: 10.1073/pnas.93.24.13677.

DOI:10.1073/pnas.93.24.13677
PMID:8942993
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC19388/
Abstract

Fidelity of DNA and protein synthesis is regulated by a proofreading mechanism but function of a similar mechanism during RNA synthesis has not been demonstrated. Analysis of transcriptional fidelity and its control has been hampered by the necessity to employ complex DNA templates requiring either a promoter and initiation factors or 3'-extended templates. To circumvent this difficulty, we have created an RNA-DNA dumbbell template that can be recognized as a template-primer and extended by RNA polymerase II. By employing this system, we demonstrate that RNA polymerase II can misincorporate a nucleotide and carry out template-dependent elongation at the mispaired end. The transcripts containing misincorporated residues can be cleaved by the very slow 3'-->5' ribonuclease activity of the RNA polymerase II, but enhancement of this activity by the elongation factor TFIIS generates RNA with a high degree of fidelity. This enhanced preferential cleavage of misincorporated transcripts suggests an important role for TFIIS in maintaining transcriptional fidelity.

摘要

DNA和蛋白质合成的保真度由校对机制调控，但RNA合成过程中类似机制的功能尚未得到证实。转录保真度及其控制的分析一直受到阻碍，因为需要使用复杂的DNA模板，这些模板要么需要启动子和起始因子，要么需要3'端延伸的模板。为了克服这一困难，我们创建了一种RNA-DNA哑铃模板，它可以被识别为模板引物，并由RNA聚合酶II进行延伸。通过使用该系统，我们证明RNA聚合酶II可以错误掺入一个核苷酸，并在错配末端进行模板依赖性延伸。含有错误掺入残基的转录本可以被RNA聚合酶II非常缓慢的3'→5'核糖核酸酶活性切割，但延伸因子TFIIS增强这种活性会产生高度保真的RNA。错误掺入转录本的这种增强的优先切割表明TFIIS在维持转录保真度方面具有重要作用。

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本文引用的文献

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The increment of SII-facilitated transcript cleavage varies dramatically between elongation competent and incompetent RNA polymerase II ternary complexes.SII促进的转录本切割的增量在具有延伸能力和不具有延伸能力的RNA聚合酶II三元复合物之间差异巨大。
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Transcript cleavage factors from E. coli.来自大肠杆菌的转录物切割因子。
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Molecular cloning of an essential subunit of RNA polymerase II elongation factor SIII.RNA聚合酶II延伸因子SIII必需亚基的分子克隆
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The transcription factor TFIIS zinc ribbon dipeptide Asp-Glu is critical for stimulation of elongation and RNA cleavage by RNA polymerase II.转录因子TFIIS锌带二肽天冬氨酸-谷氨酸对于RNA聚合酶II刺激延伸和RNA切割至关重要。
Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):9106-10. doi: 10.1073/pnas.91.19.9106.
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Conformational coupling in DNA polymerase fidelity.DNA聚合酶保真度中的构象偶联。
Annu Rev Biochem. 1993;62:685-713. doi: 10.1146/annurev.bi.62.070193.003345.
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RNA polymerase marching backward.RNA聚合酶向后行进。
Science. 1993 Feb 12;259(5097):944-5. doi: 10.1126/science.7679800.

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