Structural and biochemical analysis of DNA lesion-induced RNA polymerase II arrest

被引:6
|
作者
Oh, Juntaek [1 ]
Xu, Jun [1 ]
Chong, Jenny [1 ]
Wang, Dong [1 ,2 ]
机构
[1] Univ Calif San Diego, Skaggs Sch Pharm & Pharmaceut Sci, Div Pharmaceut Sci, La Jolla, CA 92093 USA
[2] Univ Calif San Diego, Sch Med, Dept Cellular & Mol Med, La Jolla, CA 92093 USA
基金
美国国家卫生研究院;
关键词
RNA polymerase II; X-ray crystallography; DNA lesion; Transcription; TRANSCRIPTIONAL FIDELITY; SUBSTRATE-SPECIFICITY; MOLECULAR-BASIS; TRIGGER LOOP; INITIATION; MECHANISM; ARCHITECTURE; RECOGNITION; INHIBITION; ELONGATION;
D O I
10.1016/j.ymeth.2019.02.019
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Transcription, catalyzed by RNA polymerase II (Pol II) in eukaryotes, is the first step in gene expression. RNA Pol II is a 12-subunit enzyme complex regulated by many different transcription factors during transcription initiation, elongation, and termination. During elongation, Pol II encounters various types of obstacles that can cause transcriptional pausing and arrest. Through decades of research on transcriptional pausing, it is widely known that Pol II can distinguish between different types of obstacles by its active site. A major class of obstacles is DNA lesions. While some DNA lesions can cause transient transcriptional pausing, which can be bypassed by Pol II itself or with the help from other elongation factors, bulky DNA damage can cause prolonged transcriptional pausing and arrest, which signals for transcription coupled repair. Using biochemical and structural biology approaches, the outcomes of many different types of DNA lesions, DNA modifications, and DNA binding molecules to transcription were studied. In this mini review, we will describe the in vitro transcription assays with Pol II to investigate the impacts of various DNA lesions on transcriptional outcomes and the crystallization method of lesion-arrested Pol II complex. These methods can provide a general platform for the structural and biochemical analysis of Pol II transcriptional pausing and bypass mechanisms.
引用
收藏
页码:29 / 34
页数:6
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