Mouse methyltransferase for repair of O6-methylguanine and O4-methylthymine in DNA

Hisaya Kawate, Kenji Ihara, Kohfuku Kohda, Kunihiko Sakumi, Mutsuo Sekiguchi

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

cDNA for mouse O6-methylguanine-DNA methyltransfer-ase was expressed in methyltransferase-deficient Escher-ichia coli mutant cells, and the overproduced mouse enzyme was purified to a homogeneous state. Using this purified product, polyclonal antibodies were prepared and used to estimate amounts of the methyltransferase protein in cells. A single cell of NIH3T3 contained 1.8 ×104molecules of the methyltransferase protein. When mouse fibroblasts were immunostained, it was shown that most of the methyltransferase protein exists in the cytoplasm rather than in the nucleus. Using double-stranded oligomers containing a single O6-methylguanine or O4-methylthymine at predetermined sites, the mouse enzyme repaired O6-methylguanine and O4-methylthymine, at an almost equal efficiency. In the LacZ reversion assay, MNNG-induced A: T to G: C as well as G: C to A: T transition mutations were efficiently suppressed by the function of mouse methyltransferase, in vivo.

Original languageEnglish
Pages (from-to)1595-1602
Number of pages8
JournalCarcinogenesis
Volume16
Issue number7
DOIs
Publication statusPublished - Jul 1 1995

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Methyltransferases
Protein Methyltransferases
DNA
Methylnitronitrosoguanidine
Enzymes
Cytoplasm
Complementary DNA
Fibroblasts
O-(6)-methylguanine
Mutation
Antibodies

All Science Journal Classification (ASJC) codes

  • Cancer Research

Cite this

Mouse methyltransferase for repair of O6-methylguanine and O4-methylthymine in DNA. / Kawate, Hisaya; Ihara, Kenji; Kohda, Kohfuku; Sakumi, Kunihiko; Sekiguchi, Mutsuo.

In: Carcinogenesis, Vol. 16, No. 7, 01.07.1995, p. 1595-1602.

Research output: Contribution to journalArticle

Kawate, Hisaya ; Ihara, Kenji ; Kohda, Kohfuku ; Sakumi, Kunihiko ; Sekiguchi, Mutsuo. / Mouse methyltransferase for repair of O6-methylguanine and O4-methylthymine in DNA. In: Carcinogenesis. 1995 ; Vol. 16, No. 7. pp. 1595-1602.
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