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酿酒酵母Ty1反转录转座子的转座作用会因不当的冷冻保存而被激活。

Transposition of Saccharomyces cerevisiae Ty1 retrotransposon is activated by improper cryopreservation.

作者信息

Stamenova R, Dimitrov M, Stoycheva T, Pesheva M, Venkov P, Tsvetkov T S

机构信息

Institute of Cryobiology and Food Technology, 53A "Cherni Vrah" Boulevard, 1407 Sofia, Bulgaria.

出版信息

Cryobiology. 2008 Jun;56(3):241-7. doi: 10.1016/j.cryobiol.2008.03.007. Epub 2008 Apr 7.

Abstract

Ty1 is a retrotransposon of the yeast Saccharomyces cerevisiae whose transposition at new locations in the host genome is activated by stress conditions, such as exposure to UV light, X-rays, nitrogen starvation. In this communication, we supply evidence that cooling for 2 h at +4 degrees C followed by freezing for 1 h at -10 degrees C and 16 h at -20 degrees C also increased Ty1 transposition. The mobility of Ty1 was induced by cooling at slow rates (3 degrees C/min) and the accumulation of trehalose inside cells or the cooling at high rates (100 degrees C/min) inhibited significantly the induction of the transposition. The freeze-induced Ty1 transposition did not occur in mitochondrial mutants (rho-) and in cells with disrupted SCO1 gene (Deltasco1 cells) evidencing that the Ty1 transposition induced by cooling depends on the mitochondrial oxidative phosphorylation. We also found that the freeze induced Ty1 transposition is associated with increased synthesis and accumulation of superoxide anions (O2-) into the cells. Accumulation of O2- and activation of Ty1 transposition were not observed after cooling of cells with compromised mitochondrial functions (rho-, Deltasco1), or in cells pretreated with O2- scavengers. It is concluded that (i) elevated levels of reactive oxygen species (ROS) have a key role in activation the transposition of Ty1 retrotransposon in yeast cells undergoing freezing and (ii) given the deleterious effect of increased ROS levels on cells, special precautions should be taken to avoid ROS production and accumulation during cryopreservation procedures.

摘要

Ty1是酿酒酵母中的一种逆转座子,其在宿主基因组新位置的转座可被应激条件激活,如暴露于紫外线、X射线、氮饥饿。在本通讯中,我们提供证据表明,在4℃冷却2小时,随后在-10℃冷冻1小时和在-20℃冷冻16小时也会增加Ty1转座。Ty1的移动性通过缓慢冷却速率(3℃/分钟)诱导,细胞内海藻糖的积累或快速冷却速率(100℃/分钟)显著抑制转座诱导。冷冻诱导的Ty1转座在线粒体突变体(rho-)和SCO1基因破坏的细胞(Deltasco1细胞)中不发生,这证明冷却诱导的Ty1转座依赖于线粒体氧化磷酸化。我们还发现,冷冻诱导的Ty1转座与细胞中超氧阴离子(O2-)的合成和积累增加有关。在用线粒体功能受损的细胞(rho-,Deltasco1)冷却后,或在用O2-清除剂预处理的细胞中,未观察到O2-的积累和Ty1转座的激活。得出的结论是:(i)活性氧(ROS)水平升高在经历冷冻的酵母细胞中激活Ty1逆转座子的转座中起关键作用;(ii)鉴于ROS水平升高对细胞的有害影响,在冷冻保存程序中应采取特殊预防措施以避免ROS的产生和积累。

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