Mechanisms of backtrack recovery by RNA polymerases i and II

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Ana Lisica - , Chair of Biophysics, Max Planck Institute of Molecular Cell Biology and Genetics (Author)
  • Christoph Engel - , Max Planck Institute for Biophysical Chemistry (Karl Friedrich Bonhoeffer Institute) (Author)
  • Marcus Jahnel - , Chair of Biophysics, Max Planck Institute of Molecular Cell Biology and Genetics (Author)
  • Édgar Roldán - , Max-Planck-Institute for the Physics of Complex Systems, TUD Dresden University of Technology, Universidad Carlos III de Madrid (Author)
  • Eric A. Galburt - , Washington University St. Louis (Author)
  • Patrick Cramer - , Max Planck Institute for Biophysical Chemistry (Karl Friedrich Bonhoeffer Institute) (Author)
  • Stephan W. Grill - , Chair of Biophysics, Max Planck Institute of Molecular Cell Biology and Genetics, Max-Planck-Institute for the Physics of Complex Systems (Author)

Abstract

During DNA transcription, RNA polymerases often adopt inactive backtracked states. Recovery from backtracks can occur by 1D diffusion or cleavage of backtracked RNA, but how polymerases make this choice is unknown. Here, we use single-molecule optical tweezers experiments and stochastic theory to show that the choice of a backtrack recovery mechanism is determined by a kinetic competition between 1D diffusion and RNA cleavage. Notably, RNA polymerase I (Pol I) and Pol II recover from shallow backtracks by 1D diffusion, use RNA cleavage to recover from intermediary depths, and are unable to recover from extensive backtracks. Furthermore, Pol I and Pol II use distinct mechanisms to avoid nonrecoverable backtracking. Pol I is protected by its subunit A12.2, which decreases the rate of 1D diffusion and enables transcript cleavage up to 20 nt. In contrast, Pol II is fully protected through association with the cleavage stimulatory factor TFIIS, which enables rapid recovery from any depth by RNA cleavage. Taken together, we identify distinct backtrack recovery strategies of Pol I and Pol II, shedding light on the evolution of cellular functions of these key enzymes.

Details

Original languageEnglish
Pages (from-to)2946-2951
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume113
Issue number11
Publication statusPublished - 15 Mar 2016
Peer-reviewedYes

External IDs

PubMed 26929337

Keywords

ASJC Scopus subject areas

Keywords

  • Backtracking, Optical tweezers, Pol I, Pol II, Transcription