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Genetic Analysis of Desiccation Tolerance in Saccharomyces cerevisiae

机译:酿酒酵母干燥耐性的遗传分析

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摘要

Desiccation tolerance, the ability to survive nearly total dehydration, is a rare strategy for survival and reproduction observed in all taxa. However, the mechanism and regulation of this phenomenon are poorly understood. Correlations between desiccation tolerance and potential effectors have been reported in many species, but their physiological significance has not been established in vivo. Although the budding yeast Saccharomyces cerevisiae exhibits extreme desiccation tolerance, its usefulness has been hampered by an inability to reduce tolerance more than a few fold by physiological or genetic perturbations. Here we report that fewer than one in a million yeast cells from low-density logarithmic cultures survive desiccation, while 20–40% of cells from saturated cultures survive. Using this greatly expanded metric, we show that mutants defective in trehalose biosynthesis, hydrophilins, responses to hyperosmolarity, and hypersalinity, reactive oxygen species (ROS) scavenging and DNA damage repair nevertheless retain wild-type levels of desiccation tolerance, suggesting that this trait involves a unique constellation of stress factors. A genome-wide screen for mutants that render stationary cells as sensitive as log phase cells identifies only mutations that block respiration. Respiration as a prerequisite for acquiring desiccation tolerance is corroborated by respiration inhibition and by growth on nonfermentable carbon sources. Suppressors bypassing the respiration requirement for desiccation tolerance reveal at least two pathways, one of which, involving the Mediator transcription complex, is associated with the shift from fermentative to respiratory metabolism. Further study of these regulators and their targets should provide important clues to the sensors and effectors of desiccation tolerance.
机译:耐干燥性,即几乎完全脱水的生存能力,是在所有类群中观察到的罕见的生存和繁殖策略。但是,对该现象的机理和调节知之甚少。在许多物种中已经报道了干燥耐受性与潜在效应子之间的相关性,但是它们的生理学意义尚未在体内得到证实。尽管发芽的酵母酿酒酵母显示出极高的干燥耐受性,但由于生理或遗传扰动无法将耐受性降低几倍以上,因此其实用性受到了阻碍。在这里我们报告说,低密度对数培养物中的百万个酵母细胞中只有不到一个能够存活干燥,而饱和培养物中的20-40%的细胞能够存活。使用这个大大扩展的指标,我们显示出在海藻糖生物合成,亲水蛋白,对高渗,高盐度的反应,高活性氧(ROS)清除和DNA损伤修复方面存在缺陷的突变体仍然保留了野生型水平的耐干燥性,表明该特性涉及独特的压力因素星座。在全基因组范围内筛选使固定细胞与对数期细胞一样敏感的突变体的筛选,仅能识别出阻碍呼吸作用的突变。呼吸抑制和在不可发酵碳源上的生长证实了呼吸是获得耐干燥性的前提。抑制剂绕过了对脱水耐受性的呼吸要求,显示出至少两种途径,其中一种涉及介体转录复合物,与发酵代谢向呼吸代谢的转变有关。对这些调节器及其目标的进一步研究应为干燥耐性的传感器和效应器提供重要线索。

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