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Nucleic Acids Research, 1990, Vol. 18, No. 6 1499-1508
© 1990

MOLECULAR BIOLOGY

Transcription of adenovirus and HeLa cell genes in the presence of drugs that inhibit topoisomerase I and II function

Jerome Schaak*, Paul Schedl1 and Thomas Shenk

Howard Hughes Medical Institute, Department of Biology, Princeton University Princeton, NJ 08544, USA 1Department of Biology, Princeton University Princeton, NJ 08544, USA

*To whom correspondence should be addressed

Received December 1, 1989. Revised February 11, 1990. Accepted February 11, 1990.

The requirements for topoisomerases in transcription of adenovirus and HeLa cell genes were analyzed using drugs that specifically inhibit either topoisomerase I or II. Cleavage of viral DNA by topoisomerases in the presence of either camptothecin or VM26 was used to determine drug concentrations that led to maximal inhibition of ligation in the cleavage and ligation step of topoisomerase I or II respectively. Inhibition of topoisomerase II with VM26 did not cause a direct reduction in transcription of adenoviral genes or HeLa cell heat shock genes. VM26 did, however, interfere with other cellular processes. It reduced nucleoside uptake into HeLa cells from the medium, and It altered the normal nuclear to cytoplasmic ratio of specific RNAs. Treatment of cells with camptothecin to inhibit topoisomerase I reduced but did not abolish transcription of viral and HeLa cell genes. Transcription mediated by both RNA polymerases I and II was reduced. Topoisomerase II did not appear to substitute for topoisomerase I in transcription since treatment of cells with VM26 and camptothecin did not reduce transcript accumulation relative to cells treated with camptothecin alone.


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