Generation and Elimination of 8-Oxo-7, 8-Dihydro-2'-Deoxyguanosine 5'-Triphosphate, A Mutagenic Substrate for DNA Synthesis, in Human Cells

Hiroshi Hayakawa, Michihiko Kuwano, Akinobu Taketomi, Sakumi Kunihiko, Mutsuo Sekiguchi

Research output: Contribution to journalArticle

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Abstract

8-Oxo-7, 8-dihydro-2'-deoxyguanosine 5'-triphosphate (8-oxo-dGTP) is a potent mutagenic substrate for DNA synthesis. The present study deals with generation and degradation of 8-oxo-dGTP in the nucleotide pool of human cells. (1) 8-Oxo-dGTP can be generated not only by direct oxidation of dGTP but also by phosphorylation of 8-oxo-dGDP by nucleoside diphosphate kinase. (2) 8-Oxo-dGTP is rapidly degraded to 8-oxo-dGMP by cellular 8-oxo-dGTPase activity. 8-Oxo-dGMP thus produced cannot be rephosphorylated; guanylate kinase, which phosphorylates both GMP and dGMP to the corresponding nucleoside diphosphates, is totally inactive for 8-oxo-dGMP. (3) 8-Oxo-dGMP is further degraded to 8-oxo-deoxyguanosine by a nucleotidase. The enzyme was partially purified from an extract of human Jurkat cells, and the mode of action was elucidated. 8-Oxo-dGMP is the most preferred substrate of the enzyme, and other nucleoside monophosphates are cleaved at significantly lower rates: Km for 8-oxo-dGMP is 10 times lower than that for dGMP, the second best substrate for the enzyme. The enzyme appears to convert 8-oxo-dGMP, which accumulates in the cellular nucleotide pool, to a form readily excretable to the cell exterior.

Original languageEnglish
Pages (from-to)89-95
Number of pages7
JournalBiochemistry
Volume34
Issue number1
DOIs
Publication statusPublished - Jan 1 1995

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Cells
DNA
Substrates
nucleotidase
Enzymes
Nucleosides
Nucleotides
Guanylate Kinases
Nucleoside-Diphosphate Kinase
Phosphorylation
Deoxyguanosine
Jurkat Cells
Diphosphates
8-oxo-7,8-dihydro-2'-deoxyguanosine-5'-monophosphate
8-oxodeoxyguanosine triphosphate
Degradation
Oxidation
2'-deoxyguanosine 5'-phosphate

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Generation and Elimination of 8-Oxo-7, 8-Dihydro-2'-Deoxyguanosine 5'-Triphosphate, A Mutagenic Substrate for DNA Synthesis, in Human Cells. / Hayakawa, Hiroshi; Kuwano, Michihiko; Taketomi, Akinobu; Kunihiko, Sakumi; Sekiguchi, Mutsuo.

In: Biochemistry, Vol. 34, No. 1, 01.01.1995, p. 89-95.

Research output: Contribution to journalArticle

Hayakawa, Hiroshi ; Kuwano, Michihiko ; Taketomi, Akinobu ; Kunihiko, Sakumi ; Sekiguchi, Mutsuo. / Generation and Elimination of 8-Oxo-7, 8-Dihydro-2'-Deoxyguanosine 5'-Triphosphate, A Mutagenic Substrate for DNA Synthesis, in Human Cells. In: Biochemistry. 1995 ; Vol. 34, No. 1. pp. 89-95.
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