This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Lodge, J K Articles by Berg, D E Search for Related Content PubMed PubMed Citation Articles by Lodge, J K Articles by Berg, D E

 Previous Article  |  Next Article 

J Bacteriol. 1989 April; 171(4): 2181-2187

research-article

Formation of supercoiling domains in plasmid pBR322.

Department of Microbiology and Immunology, Washington University Medical School, St. Louis, Missouri 63110.

ABSTRACT

Twin domains of positive and negative supercoiling are thought to form in DNA molecules whenever free rotation of a transcription complex around the DNA helix is impeded. Evidence for these domains has come from findings with Escherichia coli strains that are deficient in DNA topoisomerase I (top mutants) or that have been treated with DNA gyrase inhibitors. Plasmid pBR322 is highly supercoiled in these strains, whereas some of its deletion derivatives are not. The studies of pBR322 derivatives presented here show that high negative supercoiling in top strains requires translation as well as transcription of the first 98 codons of the tet gene and does not require the divergently transcribed amp gene. The N-terminal region of the TetA protein is thought to insert into the inner membrane. Our results favor models in which supercoiling domains are created when DNA segments are anchored to a large cellular structure via coupled transcription, translation, and membrane insertion of a nascent protein.

This article has been cited by other articles:

• Stupina, V. A., Wang, J. C. (2004). DNA axial rotation and the merge of oppositely supercoiled DNA domains in Escherichia coli: Effects of DNA bends. Proc. Natl. Acad. Sci. USA 101: 8608-8613 [Abstract] [Full Text]  
• Deng, S., Stein, R. A., Higgins, N. P. (2004). Transcription-induced barriers to supercoil diffusion in the Salmonella typhimurium chromosome. Proc. Natl. Acad. Sci. USA 101: 3398-3403 [Abstract] [Full Text]  
• Reyes-Dominguez, Y., Contreras-Ferrat, G., Ramirez-Santos, J., Membrillo-Hernandez, J., Gomez-Eichelmann, M. C. (2003). Plasmid DNA Supercoiling and Gyrase Activity in Escherichia coli Wild-Type and rpoS Stationary-Phase Cells. J. Bacteriol. 185: 1097-1100 [Abstract] [Full Text]  
• Bentin, T., Nielsen, P. E. (2002). In vitro transcription of a torsionally constrained template. Nucleic Acids Res 30: 803-809 [Abstract] [Full Text]  
• Peng, H. F., Jackson, V. (2000). In Vitro Studies on the Maintenance of Transcription-induced Stress by Histones and Polyamines. J. Biol. Chem. 275: 657-668 [Abstract] [Full Text]  
• D'Argenio, D. A., Segura, A., Coco, W. M., Bünz, P. V., Ornston, L. N. (1999). The Physiological Contribution of Acinetobacter PcaK, a Transport System That Acts upon Protocatechuate, Can Be Masked by the Overlapping Specificity of VanK. J. Bacteriol. 181: 3505-3515 [Abstract] [Full Text]  
• Chen, D., Bachellier, S., Lilley, D. M. J. (1998). Activation of the leu-500 Promoter by a Reversed Polarity tetA Gene. RESPONSE TO GLOBAL PLASMID SUPERCOILING. J. Biol. Chem. 273: 653-659 [Abstract] [Full Text]  
• Ostrander, E., Benedetti, P, Wang, J. (1990). Template supercoiling by a chimera of yeast GAL4 protein and phage T7 RNA polymerase. Science 249: 1261-1265 [Abstract]  
• Rahmouni, A., Wells, R. (1989). Stabilization of Z DNA in vivo by localized supercoiling. Science 246: 358-363 [Abstract]