TY - JOUR

T1 - Evidence that Transcript Cleavage Is Essential for RNA Polymerase II Transcription and Cell Viability

AU - Sigurdsson, Stefan

AU - Dirac-Svejstrup, A. Barbara

AU - Svejstrup, Jesper Q.

N1 - 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.

PY - 2010

Y1 - 2010

N2 - 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.

AB - 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.

KW - DNA

KW - RNA

U2 - 10.1016/j.molcel.2010.02.026

DO - 10.1016/j.molcel.2010.02.026

M3 - Journal article

C2 - 20417599

AN - SCOPUS:77950998789

VL - 38

SP - 202

EP - 210

JO - Molecular Cell

JF - Molecular Cell

SN - 1097-2765

IS - 2

ER -