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Ultrasensitive tracking of oxidized guanine incorporation into DNA and RNA via nucleotide salvage with accelerator mass spectrometry

등록일
2009년 8월 5일 14시 42분 35초
접수번호
0061
발표코드
금15H6심 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
발표시간
금 12시 : 00분
발표형식
심포지엄
발표분야
생명화학 - Nanomaterials and Nucleic Acids for Molecular Therapy
저자 및
공동저자
하상수
경희대학교 화학과, Korea
Growing evidence suggests that oxidative damage to cells generates mutagenic 7,8-dihydro-8-oxo-2’-deoxyguanosine (8-oxodG), which initiates diseases related to aging and carcinogenesis. Insufficient detection sensitivity, however, has hampered measurement of these putative mutagenic products in cells or animals. I report use of a sensitive and precise accelerator mass spectrometry (AMS) assay to track the fate of [14C]8-oxodG incorporation into DNA and RNA of MCF-7 human breast cancer cells. MCF-7 cells were grown in the presence of [14C]8-oxodG and radiocarbon was found to be partitioned into both DNA and RNA, indicating that 8-oxoguanine was detached from the deoxyribose ring and attached covalently to a ribosyl species to form an appropriate nucleotide for incorporation into RNA. I also observed evidence for phosphorylated 8-oxodG derivatives and 8-oxoG derivatives in the nucleotide pool. Moreover, oxidative stressing of the cells by exposure to 17ß-estradiol reduced the rate of 8-oxodG incorporation, indicating induction of a mechanism to limit the triphosphate derivative of 8-oxodG required for incorporation by DNA polymerases. These data suggest that 8-oxodG in the nucleotide pool is an unexpected source of 8-oxoguanine forDNA and RNA incorporation, which may have important consequences for understanding diseases resulting from mutagenesis and altered protein function.

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