Abstract
During transcript elongation in vitro, backtracking of RNA polymerase II (RNAPII) is a frequent occurrence that can lead to transcriptional arrest. The polymerase active site can cleave the transcript during such backtracking, allowing transcription to resume. Transcript cleavage is either stimulated by elongation factor TFIIS or occurs much more slowly in its absence. However, whether backtracking actually occurs in vivo, and whether transcript cleavage is important to escape it, has been unclear. Using a yeast TFIIS mutant that lacks transcript cleavage stimulatory activity and simultaneously inhibits unstimulated cleavage, we now provide evidence that escape from backtracking via transcript cleavage is essential for cell viability and efficient transcript elongation. Our results suggest that transcription problems leading to backtracking are frequent in vivo and that reactivation of backtracked RNAPII is crucial for transcription.
| Original language | English |
|---|---|
| Pages (from-to) | 202-210 |
| Number of pages | 9 |
| Journal | Molecular Cell |
| Volume | 38 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 23 Apr 2010 |
Bibliographical note
Funding Information: This work was supported by an EMBO long-term postdoctoral fellowship to S.S., and by grants from Cancer Research UK, Association of International Cancer Research, and the European Community (Integrated Project DNA repair, LSHG-CT-2005-512113) to J.Q.S. Caroline Kane is thanked for the His-TFIIS expression plasmid; David Jansma and Jim Friesen are thanked for yeast strains; and Dong Wang, Dave Bushnell, Craig Kaplan, and Roger Kornberg are thanked for helpful discussions and advice on RNAPII structure, as well as for comments on the manuscript. Peter Verrijzer and members of the Svejstrup laboratory are thanked for discussions and helpful comments on the manuscript.Other keywords
- DNA
- RNA