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Synthesis of Prematurely Terminated Late Transcripts of Polyoma Virus DNA is Resistant to Inhibition by 5,6-Dichloro-1--D-ribofuranosylbenzimidazole

Paul-Etienne Montandon

Nicholas H. Acheson

Swiss Institute for Experimental Cancer Research, 1066 Epalinges, Switzerland

Exposure of mouse kidney cells to 5,6–dichloro–1–-D–ribofuranosylbenzimidazole (DRB) (75 to 300 µM) during the late phase of infection by polyoma virus resulted in nearly complete (90 to 98%) inhibition of virus RNA synthesis. Sedimentation analysis revealed that, although the synthesis of high mol. wt. (> 10S) virus RNA was inhibited in a manner parallel to that of total virus RNA, the synthesis of small (3S to 7S) virus RNA was inhibited by only 40 to 50% in the presence of DRB. As a result, virus RNA synthesized in the presence of DRB contained a peak at 3S to 7S in addition to residual high mol. wt. virus RNA. Small virus RNA from either untreated or DRB-treated cells contained three- to sixfold higher levels of transcripts from the DNA fragment which lies between the BamHI and BglI sites (58.0 to 72.2 map units) than from DNA fragments covering the rest of the virus genome. Furthermore, 80% of the small RNA which hybridized to this fragment was complementary to the L strand of virus DNA. These results suggest that L strand transcripts are initiated within the BglI–BamHI DNA fragment and that a portion of these transcripts is prematurely terminated within several hundred nucleotides of the site(s) of initiation. DRB had little effect on the synthesis of these prematurely terminated RNAs.

Keywords: DNA tumour virus, transcription, synthesis, inhibition

Present address: Tumor Virology Laboratory, The Salk Institute, San Diego, California 92138, U.S.A.

Present address: Department of Microbiology, McGill University, Montreal, Quebec H3A 2B4, Canada.

Received 6 July 1981; accepted 12 October 1981.