Induction of DNA double-strand breaks by zeocin in Chlamydomonas reinhardtii and the role of increased DNA double-strand breaks rejoining in the formation of an adaptive response

Abstract

This study aimed to test the potential of the radiomimetic chemical zeocin to induce DNA double-strand breaks (DSB) and "adaptive response" (AR) in Chlamydomonas reinhardtii strain CW15 as a model system. The AR was measured as cell survival using a micro-colony assay, and by changes in rejoining of DSB DNA. The level of induced DSB was measured by constant field gel electrophoresis based on incorporation of cells into agarose blocks before cell lysis. This avoids the risk of accidental induction of DSB during the manipulation procedures. Our results showed that zeocin could induce DSB in C. reinhardtii strain CW15 in a linear dose-response fashion up to 100 microg ml(-1) which marked the beginning of a plateau. The level of DSB induced by 100 microg ml(-1) zeocin was similar to that induced by 250 Gy of gamma-ray irradiation. It was also found that, similar to gamma rays, zeocin could induce AR measured as DSB in C. reinhardtii CW15 and this AR involved acceleration of the rate of DSB rejoining, too. To our knowledge, this is the first demonstration that zeocin could induce AR in some low eukaryotes such as C. reinhardtii.