First Author | Kawate H | Year | 1995 |
Journal | Carcinogenesis | Volume | 16 |
Issue | 7 | Pages | 1595-602 |
PubMed ID | 7614694 | Mgi Jnum | J:26835 |
Mgi Id | MGI:74265 | Doi | 10.1093/carcin/16.7.1595 |
Citation | Kawate H, et al. (1995) Mouse methyltransferase for repair of O6-methylguanine and O4-methylthymine in DNA. Carcinogenesis 16(7):1595-602 |
abstractText | cDNA for mouse O6-methylguanine-DNA methyltransferase was expressed in methyltransferase-deficient Escherichia 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 x 10(4) molecules 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. |