Nucleic Acids Research, 1992, Vol. 20, No. 22 5991-5997
© 1992
MOLECULAR BIOLOGY |
Transcriptionally driven cruciform formation in vivo
Department of Genetics, University of Illinois at Chicago 808 S.Wood Street, Chicago, IL 60612 1California Institute of Biological Research La Jolla, CA 92037, USA
* To whom correspondence should be addressed
Received August 13, 1992. Revised October 25, 1992. Accepted October 25, 1992.
We studied the formation of d(A-T)n cruciforms in E.coli cells by probing intracellular plasmid DNA with chloroacetaldehyde followed by fine analysis of modified DNA bases. d(A-T)16 sequences were inserted into specifically designed plasmids either upstream of a single trc promoter, or between two divergent trc promoters. We found that in both cases, induction of transcription by IPTG leads to the transition of the d(A-T)16 stretch into a cruciform state. In the case of two divergent promoters, we observed cruciform formation even without IPTG. Enhanced cruciform formation correlates with the elevation in promoter activity as defined by the opening of the promoter at the 
10 to +2 positions. We conclude that transcriptionally driven negative supercoiling provokes cruciform formation in vivo
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