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Isolation and characterization of mutator mutants from cultured mouse FM3A cells
Authors:Masao Hyodo  Narushi Ito  Hideki Koyama  Kenshi Suzuki
Institution:

* Department of Molecular Biology, Tokai University School of Medicine, Boseidai, Isehara, Kanagawa, 259-11, Japan

a Department of Biochemistry, Cancer Institute, Japanese Foundation for Cancer Research, 1-37-1, Kami-Ikeburuko, Toshima, Tokyo 170, Japan

Abstract:A method to select mutator mutants was developed and 3 mutants were isolated from cultured mouse FM3A cells. Fluctuation analyses revealed that these mutator mutants have increased rates of spontaneous mutation at 3 genetic loci tested (resistance to ouabain, blasticidin S and tunicamycin). None of the 3 mutator mutants showed altered sensitivity to aphidicolin or arabinofuranosylcytosine, and so they differed from the mammalian mutator mutants reported previously. Also, all the mutator mutants had the same sensitivity as wild-type to UV or other DNA-damaging agents. Thus, these mutator mutants do not seem to have any deficiency in the DNA-repair process.

To determine whether the mutator activity was due to the intracellular dNTP pool imbalance, 4 dNTPs in these mutator mutants were determined by high-pressure liquid chromatography and compared to that of the wild-type cells. The results show that there is no large dNTP pool imbalance in these mutator mutants. Since the mutator activity is not associated with the dNTP pool imbalance, these mutants may have altered protein(s) directly involved in DNA replication.

Keywords:All correspondence should be addressed to: Masao Hyodo  Department of Molecular Biology  Tokai University School of Medicine  Boseidai  Isehara  Kanagawa 259-11 Japan
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