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/sup +/ DNAs from a variety of mammalian cell lines permitted the selection of TS/sup +/ colonies at 40/sup 0/C. TS/sup +/ transformation events were distinguished from spontaneous TS/sup +/ reversions in experiments in which ..cap alpha..-amanitin-sensitive (Ama/sup s/) TS/sup +/ DNA was used to transform an AMA/sup R/ derivative of TsAF8 cells and Ama/sup R/ TS/sup +/ DNA was used to transform Ama/sup s/ TsAF8 cells. In each case it was possible to demonstrate the unselected acquisition of the appropriate Ama/sup s/ or Ama/sup R/ phenotype with the selected TS/sup +/ allele. Each of these TS/sup +/ transformed cell lines when grown at 40/sup 0/C contained an RNA polymerase II activity with a sensitivity to inhibition by ..cap alpha..-amanitin characteristic of the particular DNA used to transform the TS cells, whereas at 34/sup 0/C the same cells contained a mixture of Ama/sup R/ and Ama/sup s/ polymerase II activities. Together, these data provide convincing evidence that the RNA polymerase II gene determining sensitivity to inhibition by ..cap alpha..-amanitin can be transferred to TsAF8 cells and that the TS defect in TsAF8 is a polymerase II mutation.
- Research Organization:
- Banting and Best Dept. of Medical Research, Univ. of Toronto, Toronto, Ontario M5G 1L6
- OSTI ID:
- 5809796
- Journal Information:
- Mol. Cell. Biol.; (United States), Vol. 2:6
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
DNA
SENSITIVITY
HAMSTERS
GENOME MUTATIONS
REVERTANTS
TEMPERATURE EFFECTS
RNA POLYMERASES
BIOLOGICAL FUNCTIONS
ENZYME ACTIVITY
ENZYME INHIBITORS
GENES
CELL CULTURES
CELL TRANSFORMATIONS
CHEMICAL REACTION KINETICS
PHENOTYPE
ANIMALS
ENZYMES
FUNCTIONS
KINETICS
MAMMALS
MUTANTS
MUTATIONS
NUCLEIC ACIDS
NUCLEOTIDYLTRANSFERASES
ORGANIC COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
REACTION KINETICS
RODENTS
TRANSFERASES
VERTEBRATES
550400* - Genetics
550200 - Biochemistry