Identification of RCN1 and RSA3 as ethanol-tolerant genes in Saccharomyces cerevisiae using a high copy barcoded library
Identification of RCN1 and RSA3 as ethanol-tolerant genes in Saccharomyces cerevisiae using a high copy barcoded library
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DOI：
10.1111/j.1567-1364.2011.00762.x
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发表时间：
2012-02-01
影响因子：
3.2
通讯作者：
Measday, Vivien
中科院分区：
文献类型：
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作者：
Anderson, Michael J.;Barker, Sarah L.;Measday, Vivien
酿酒酵母（Cerevisiae）在葡萄酒发酵等工业过程中遇到多种应力，包括乙醇毒性。高水平的乙醇降低了酵母的生存能力，并可能阻止发酵完成。耐乙醇基因的鉴定对于创建耐心的工业酵母很重要，并且用于此目的的伯晶基因组资源已用于此目的。我们采用了分子条形码酵母开放式阅读框（MOBY-ORF）高拷贝质粒文库，以鉴定在酿酒酵母S288C实验室和M2葡萄酒菌株中识别耐乙醇的基因。我们发现，RCN1或RSA3的剂量增加可提高S288C和M2的耐受性，对乙醇的毒性水平。 RCN1是钙调蛋白的调节剂，而RSA3在核糖体成熟中起作用。赋予RCN1和RSA3过量生产的其他适应性优势包括对细胞壁降解，热，渗透和氧化应激的抗性提高。我们发现，M2葡萄酒酵母菌菌株通常比S288C更容忍应激，但翻译抑制除外，这比S288C更严重地影响M2的生长。我们得出的结论是，调节核糖体生物发生和最终翻译是在与工业相关的环境压力期间经晶生存的关键因素。
Saccharomyces cerevisiae (S.cerevisiae) encounters a multitude of stresses during industrial processes such as wine fermentation including ethanol toxicity. High levels of ethanol reduce the viability of yeast and may prevent completion of fermentation. The identification of ethanol-tolerant genes is important for creating stress-resistant industrial yeast, and S.cerevisiae genomic resources have been utilized for this purpose. We have employed a molecular barcoded yeast open reading frame (MoBY-ORF) high copy plasmid library to identify ethanol-tolerant genes in both the S.cerevisiae S288C laboratory and M2 wine strains. We find that increased dosage of either RCN1 or RSA3 improves tolerance of S288C and M2 to toxic levels of ethanol. RCN1 is a regulator of calcineurin, whereas RSA3 has a role in ribosome maturation. Additional fitness advantages conferred upon overproduction of RCN1 and RSA3 include increased resistance to cell wall degradation, heat, osmotic and oxidative stress. We find that the M2 wine yeast strain is generally more tolerant of stress than S288C with the exception of translation inhibition, which affects M2 growth more severely than S288C. We conclude that regulation of ribosome biogenesis and ultimately translation is a critical factor for S.cerevisiae survival during industrial-related environmental stress.