Oligonucleotide transformation for the study of mutagenic specificities of DNA lesions in yeast.
We developed a method for the analyzing mutagenic potential of DNA damage based on the oligonucleotide transformation technique in yeast. Using this assay we have analyzed mutagenic specificities of various DNA lesions. In the present study, we analyzed the mutagenic properties of 2-hydroxyadenine and 5-hydroxycytosine in yeast. Oligonucleotides containing 2-hydroxyadenine or 5-hydroxycytosine were used for the transformation. The oligonucleotides showed transforming activities similar to unmodified oligonucleotides. This indicates that no repair systems were working on them. The sequencing data of the transformants showed that 5-hydroxycytosine and 2-hydroxyadenine are read mainly as cytosine and adenine. We will also discuss the mechanism of oligonucleotide transformation and its application to the mutagenesis study.
Negishi
Kazuo
K
Nihon Pharmaceutical University, 10281 Komuro, Ina, Saitama 362-0806, Japan. knegishi@nichiyaku.ac.jp
Sekine
Daijiro
D
Morimitsu
Takumi
T
Suzuki
Tetsuya
T
Okugawa
Yoji
Y
Kawakami
Asako
A
Otsuka
Chie
C
Oyama
Hiroshi
H
Loakes
David
D
D003596:Cytosine / Q000031:analogs & derivatives* / Q000737:chemistry; D004249:DNA Damage; D004252:DNA Mutational Analysis; D006147:Guanine / Q000737:chemistry*; D016296:Mutagenesis; D009841:Oligonucleotides / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014170:Transformation, Genetic
D009841:Oligonucleotides; D006147:Guanine; C017400:5-hydroxycytosine; D003596:Cytosine; C008183:isoguanine