TY - JOUR

T1 - Stability, Flexibility, and Dynamic Interactions of Colliding RNA Polymerase II Elongation Complexes

AU - Saeki, Hideaki

AU - Svejstrup, Jesper Q.

N1 - Funding Information: This work was supported by a grant from the European Community (Integrated Project DNA repair, grant number LSHG-CT-2005-512113) and by an in-house grant from Cancer Research UK (to J.Q.S.). We thank Stefan Sigurdsson for recombinant yeast TFIIS and helpful discussions and Caroline Kane for the TFIIS expression plasmid. Dave Bushnell and Roger Kornberg are thanked for sharing unpublished information and their insight into the RNAPII structure. Members of the Svejstrup lab, Dale Wigley, Craig Kaplan, Dave Bushnell, Dong Wang, Roger Kornberg, and Peter Verrijzer are thanked for comments on the manuscript.

PY - 2009/7/31

Y1 - 2009/7/31

N2 - Multiple RNA polymerase II (RNAPII) molecules can transcribe a gene simultaneously, but what happens when such polymerases collide-for example due to polymerase pausing or DNA damage? Here, RNAPII collision was characterized using a reconstituted system for simultaneous transcription by two polymerases. When progression of leading polymerase is obstructed, rear-end collision entails a transient state in which the elongation complexes interact, followed by substantial backtracking of trailing polymerase. Elongation complexes remain stable on DNA, with their activity and the integrity of transcription bubbles remaining intact. Subsequent TFIIS-stimulated transcript cleavage allows resumed forward translocation, resulting in trailing polymerase oscillating at the obstruction. Conversely, if leading polymerase is merely stalled at a pause site, collision and TFIIS cooperate to drive it through. We propose that dynamic interactions between RNAPII elongation complexes help regulate polymerase traffic and that their conformational flexibility buffers the effect of collisions with objects on DNA, thereby maintaining stability in the face of obstacles to transcription.

AB - Multiple RNA polymerase II (RNAPII) molecules can transcribe a gene simultaneously, but what happens when such polymerases collide-for example due to polymerase pausing or DNA damage? Here, RNAPII collision was characterized using a reconstituted system for simultaneous transcription by two polymerases. When progression of leading polymerase is obstructed, rear-end collision entails a transient state in which the elongation complexes interact, followed by substantial backtracking of trailing polymerase. Elongation complexes remain stable on DNA, with their activity and the integrity of transcription bubbles remaining intact. Subsequent TFIIS-stimulated transcript cleavage allows resumed forward translocation, resulting in trailing polymerase oscillating at the obstruction. Conversely, if leading polymerase is merely stalled at a pause site, collision and TFIIS cooperate to drive it through. We propose that dynamic interactions between RNAPII elongation complexes help regulate polymerase traffic and that their conformational flexibility buffers the effect of collisions with objects on DNA, thereby maintaining stability in the face of obstacles to transcription.

KW - DNA

U2 - 10.1016/j.molcel.2009.06.009

DO - 10.1016/j.molcel.2009.06.009

M3 - Journal article

C2 - 19647516

AN - SCOPUS:67651095903

VL - 35

SP - 191

EP - 205

JO - Molecular Cell

JF - Molecular Cell

SN - 1097-2765

IS - 2

ER -