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Nucleic Acids Research, 1995, Vol. 23, No. 10 1782-1789
© 1995

MOLECULAR BIOLOGY

Presence of negative torsional tension in the promoter region of the transcriptionally poised dihydrofolate reductase gene in vivo

Mats Ljungman*, and Philip C. Hanawalt1

Department of Radiation Oncology, Division of Cancer Biology, University of Michigan Medical Center 1331 East Ann Street, Ann Arbor, Ml 48109-0582, USA 1Department of Biological Sciences, Stanford University, Stanford CA 94305-5020, USA

* To whom correspondence should be addressed

Received January 17, 1995. Revised March 27, 1995. Accepted March 27, 1995.

DNA topology has been suggested to play an Important role In the process of transcription. Negative torsional tension has been shown to stimulate both pre-inltiation complex formation and promoter clearance on plasmld DNA in vitro We recently showed that genomlc DNA In human cells contains localized torsional tension. In the present study we have further characterized and mapped torsional tension in the dihydrofolate reductase (DHFR) gene In Chinese hamster ovary (CHO) cells and investigated the effects of differential rates of transcription on the magnitude and location of this tension. Using psoralen photo-cross-llnklng In conjunction with X-lrradlation, we found that relaxable psoralen hypersensltivtty was specifically localized to the promoter region of the serum-regulated DHFR gene in serum-stimulated, but not In serum-starved, cells. Moreover, this hypersensl-tivity did not appear to be caused by transcription elongation, since it persisted, In cells In which transcription of the DHFR gene had been reduced by the transcription inhibitor 5,6-dichloro-1-ß-D-ribofurano-sylbenzimidazole (DRB). We suggest that the generation of negative torsional tension in DNA may play an important role in gene regulation by poising genes for transcription.


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