DNA-mediated transfer of an RNA polymerase II gene: reversion of the temperature-sensitive hamster cell cycle mutant TsAF8 by mammalian DNA.
Treatment of the TsAF8 temperature-sensitive (TS) mutant of Syrian hamster BHK-21 cells, with calcium phosphate precipitates of genomic TS+ DNAs from a variety of mammalian cell lines permitted the selection of TS+ colonies at 40 degrees C. TS+ transformation events were distinguished from spontaneous TS+ reversions in experiments in which alpha-amanitin-sensitive (Amas) TS+ DNA was used to transform an AmaR derivative of TsAF8 cells and AmaR TS+ DNA was used to transform Amas TsAF8 cells. In each case it was possible to demonstrate the unselected acquisition of the appropriate Amas or AmaR phenotype with the selected TS+ allele. Each of these TS+ transformed cell lines when grown at 40 degrees C contained an RNA polymerase II activity with a sensitivity to inhibition by alpha-amanitin characteristic of the particular DNA used to transform the TS cells, whereas at 34 degrees C the same cells contained a mixture of AmaR and Amas polymerase II activities. Together, these data provide convincing evidence that the RNA polymerase II gene determining sensitivity to inhibition by alpha-amanitin can be transferred to TsAF8 cells and that the TS defect in TsAF8 is a polymerase II mutation.
Banting and Best Department of Medical Research, University of Toronto, Toronto, Ontario, Canada M5G 1L6.
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D000546:Amanitins / Q000494:pharmacology; D000818:Animals; D016466:CHO Cells; D002453:Cell Cycle / Q000235:genetics*; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D006224:Cricetinae; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics*; D006801:Humans; D009154:Mutation / Q000235:genetics*; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D010641:Phenotype; D012319:RNA Polymerase II / Q000378:metabolism*; D013696:Temperature; D014158:Transcription, Genetic / Q000235:genetics; D014170:Transformation, Genetic; D019076:Transgenes / Q000235:genetics*
D000546:Amanitins; D019384:Nucleic Acid Synthesis Inhibitors; D004247:DNA; D012319:RNA Polymerase II