We examined mechanisms involved in delayed mutagenesis in CHO-LacZeo cells harboring the fusion gene between the bacterial LacZ and the Zeocin-resistance genes. After X irradiation, Zeocin-resistant primary colonies were isolated, and the primary clones were subjected to the secondary colony formation in the absence of Zeocin. We found that the surviving primary clones showed a significantly higher delayed mutation frequency compared with those derived from nonirradiated CHO-LacZeo cells. The mutation spectrum of the LacZ gene was analyzed by the LacZ gene-specific PCR. We found that more than 90% of the spontaneous and direct mutants were PCR-product negative, indicating that deletion of the LacZ gene was a predominant change in these mutants. While deletion of the LacZ gene was also observed in delayed mutants, we found that more than 20% of delayed mutants had a PCR product similar to that of the parental CHO-LacZeo cells. These PCR product-positive mutants spontaneously reverted to LacZ-positive (LacZ+) cells, and all of these mutants became LacZ-positive after 5-azacytidine treatment. These results indicate that epigenetic gene silencing, in addition to elevated recombination, is involved in delayed mutagenesis, which is a novel mechanism underlying delayed manifestations of radiation-induced genomic instability.
Epigenetic Gene Silencing is a Novel Mechanism Involved in Delayed Manifestation of Radiation-Induced Genomic Instability in Mammalian Cells
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Keiji Suzuki, Hiroko Yamaji, Shinko Kobashigawa, Rie Kawauchi, Kazutaka Shima, Seiji Kodama, Masami Watanabe; Epigenetic Gene Silencing is a Novel Mechanism Involved in Delayed Manifestation of Radiation-Induced Genomic Instability in Mammalian Cells. Radiat Res 1 April 2011; 175 (4): 416–423. doi: https://doi.org/10.1667/RR2391.1
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