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Widespread Backtracking by RNA Pol II Is a Major Effector of Gene Activation, 5' Pause Release, Termination, and Transcription Elongation Rate.

Mol Cell. 2019-01; 
SheridanRyan M,FongNova,D'AlessandroAngelo,BentleyDav
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Recombinant Proteins Rabbit anti Avi-tag was purchased from GenScript (A00674). n. Each lysate was divided into two IPs using either 15 mL of rabbit anti-total pol II CTD serum or 5 ug anti-Avi-tag antibody (GenScript, A00674). Get A Quote

摘要

In addition to phosphodiester bond formation, RNA polymerase II has an RNA endonuclease activity, stimulated by TFIIS, which rescues complexes that have arrested and backtracked. How TFIIS affects transcription under normal conditions is poorly understood. We identified backtracking sites in human cells using a dominant-negative TFIIS (TFIIS) that inhibits RNA cleavage and stabilizes backtracked complexes. Backtracking is most frequent within 2?kb of start sites, consistent with slow elongation early in transcription, and in 3' flanking regions where termination is enhanced by TFIIS, suggesting that?backtracked pol II is a favorable substrate for?termination. Rescue from backtracking by RNA cleavage... More

关键词

RNA polymerase pausing,TFIIS,hypoxia response,transcription elongation,transcription termination,transcriptional backtrac