Mutagenicity of 5-bromouracil and N6-hydroxyadenine studied by yeast oligonucleotide transformation assay.
The mutagenicity of 5-bromouracil (BrU) and N6-hydroxyadenine (HA) was tested by means of the yeast oligonucleotide transformation procedure. BrU-containing oligonucleotide was not mutagenic; although two mutants (per 200 micrograms oligonucleotide) were obtained, they were attributed to base insertion or base substitution at positions different from BrU. This result supports the view that BrU mutagenesis is dependent on intracellular nucleotide pool imbalance. In contrast, HA-containing oligonucleotide was highly mutagenic; 56 mutants (per 140 micrograms oligonucleotide) were obtained. Of 21 induced mutants examined, 20 had G and one had C at the HA position, a result indicating that HA-->G changes took place. To provide back-up evidence, we carried out a general reversion assay for base HA using a set of yeast tester strains, and the results showed that HA induces exclusively AT-to-GC and GC-to-AT transitions. We conclude that in S. cerevisiae HA is a classic base analog mutagen, causing AT-to-GC and GC-to-AT transitions by ambiguous base pairing. The present work has clearly demonstrated the usefulness of the oligonucleotide transformation procedure for elucidating mutagenicity of modified bases.
Noskov
V
V
Gene Research Center, Okayama University, Japan.
Negishi
K
K
Ono
A
A
Matsuda
A
A
Ono
B
B
Hayatsu
H
H
D000225:Adenine / Q000031:analogs & derivatives* / Q000633:toxicity*; D000595:Amino Acid Sequence; D001483:Base Sequence; D001976:Bromouracil / Q000633:toxicity*; D008969:Molecular Sequence Data; D009152:Mutagenicity Tests; D009841:Oligonucleotides / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014170:Transformation, Genetic
D009841:Oligonucleotides; D001976:Bromouracil; D000225:Adenine