Coupling DNA supercoiling to transcription in defined protein systems.

The Journal of Biological Chemistry
Fenfei LengRoger McMacken

Abstract

Transcription of closed circular DNA templates in the presence of DNA gyrase is known to stimulate negative DNA supercoiling both in vivo and in vitro. It has proven elusive, however, to establish a general system in vitro that supports transcription-coupled DNA supercoiling (TCDS) by the "twin-domain" mechanism (Liu, L. F. and Wang, J. C. (1987) Proc. Natl. Acad. Sci. USA 84, 7024-7027) that operates in bacteria. In this report, we examine the properties of TCDS in defined protein systems that minimally contained T7 RNA polymerase and DNA gyrase. Specifically designed plasmid DNA templates permitted us to control the location and length of RNA transcripts. We demonstrate that TCDS takes place by two separate, and apparently independent, mechanistic pathways in vitro. The first supercoiling pathway, which is not likely to be significant in vivo, was found to be dependent on R-loop formation and could be suppressed by the presence of RNase H or bacterial HU protein. The second pathway for TCDS was much more potent, but became predominant in vitro only when sequence-specific DNA-bending proteins were present during transcription, and RNA transcript lengths exceeded 3 kb. This major supercoiling route was shown to be resistant to ...Continue Reading

References

Nov 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·M GellertH A Nash
Nov 30, 1979·Science·P O Brown, N R Cozzarelli
Nov 5, 1975·Journal of Molecular Biology·J P Richardson
Jun 11, 1991·Nucleic Acids Research·P Dröge, A Nordheim
Oct 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·L F Liu, J C Wang
Dec 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·G J Pruss, K Drlica
Oct 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·M S WoldR McMacken
Feb 1, 1994·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·P Dröge
Aug 20, 1993·Journal of Molecular Biology·L E MacdonaldW T McAllister
Jan 1, 1996·Annual Review of Biochemistry·J C Wang
Aug 1, 1996·Quarterly Reviews of Biophysics·D M LilleyR P Bowater
Jan 17, 1997·The Journal of Biological Chemistry·P PhoenixM Drolet
Aug 29, 1997·Journal of Molecular Biology·Z Wang, P Dröge
Jan 8, 1999·Journal of Molecular Biology·D Chen, D M Lilley
Mar 14, 2000·The Journal of Biological Chemistry·E L ZechiedrichN R Cozzarelli
Jul 3, 2002·Proceedings of the National Academy of Sciences of the United States of America·Fenfei Leng, Roger McMacken

❮ Previous
Next ❯

Citations

Nov 30, 2011·Proceedings of the National Academy of Sciences of the United States of America·Fenfei LengDavid D Dunlap
Sep 30, 2008·Nucleic Acids Research·Seung-Won ParkChee-Gun Lee
Oct 17, 2014·Proceedings of the National Academy of Sciences of the United States of America·Yue DingLaura Finzi
Mar 31, 2005·Current Opinion in Genetics & Development·Martin ThanbichlerLucy Shapiro
Oct 6, 2016·Biophysical Journal·Brad A Krajina, Andrew J Spakowitz
Oct 4, 2015·American Journal of Physiology. Lung Cellular and Molecular Physiology·Viktor M PastukhMark N Gillespie
Aug 15, 2020·Briefings in Bioinformatics·Zhaoqian LiuBingqiang Liu
Jul 12, 2017·The Journal of Biological Chemistry·Xiaoduo ZhiFenfei Leng
Jul 20, 2019·BMC Molecular and Cell Biology·Charles J Dorman
Dec 10, 2017·Scientific Reports·Peter GroßmannChristoph Kaleta
Jan 13, 2019·Proceedings of the National Academy of Sciences of the United States of America·Jie MaMichelle D Wang
Mar 10, 2017·Biophysics Reviews·Jie Ma, Michelle D Wang
Apr 22, 2020·Nucleic Acids Research·Omar L Kantidze, Sergey V Razin

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.