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Nucleic Acids Research, 1977, Vol. 4, No. 6 1713-1726
© 1977

Articles

Inhibition of transcription of supercoiled PM2 DNA by carbodiimide modification

Marcia S. Flashner*, Mary Ann Katopes* and Jacob Lebowitz{dagger},{dagger}

*Dep. Biol., Biol. Res. Lab., Syracuse Univ. Syracuse, NY 13210 {dagger}Dep. Microbiol., Box 11DB, and Comprehensive Cancer Cent., Univ. Alabama in Birmingham Birmingham, AL 35294, USA

Received December 15, 1976.

PM2 super-helical DNA (Form I), which has been reacted with the single strand specific reagent, N-cyclohexyl-N'-ß-(methylmorpholinium)ethyl carbodiimide (CMC) is more than 95% inhibited in its ability to support transcription with E. coli B RNA polymerase in vitro. Almost complete inhibition of transcription was achieved after 2 hours of reaction with FI when only 1% of the bases were modified. A large increase in S20,* (from 26.8 S to 33.6 S) of FI DNA was observed during the course of reaction. Rifampicin resistant transcription is more susceptible to inhibition by CMC than total transcription, suggesting that the CMC is preferentially binding at promoter sites. These results clearly are in accord with the observation that supercoiled DNA contains localized regions of unpaired bases. The promotor sites for E. coli RNA polymerase in FI PM2 DNA appear to be located at or near these unpaired sites.

{dagger}Address all reprint requests to L. Lebowitz.


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