DNA-mediated transfer of an RNA polymerase II gene: reversion of the temperature-sensitive hamster cell cycle mutant TsAF8 by mammalian DNA

Mol Cell Biol. 1982 Jun;2(6):666-73. doi: 10.1128/mcb.2.6.666-673.1982.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amanitins / pharmacology
  • Animals
  • CHO Cells
  • Cell Cycle / genetics*
  • Cell Division / drug effects
  • Cell Line
  • Cricetinae
  • DNA / administration & dosage*
  • DNA / genetics*
  • Humans
  • Mutation / genetics*
  • Nucleic Acid Synthesis Inhibitors / pharmacology
  • Phenotype
  • RNA Polymerase II / metabolism*
  • Temperature
  • Transcription, Genetic / genetics
  • Transformation, Genetic*
  • Transgenes / genetics*

Substances

  • Amanitins
  • Nucleic Acid Synthesis Inhibitors
  • DNA
  • RNA Polymerase II