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首页> 外文期刊>Comparative and functional genomics >Genome-Wide Analysis of the Effects of Heat Shock on aSaccharomyces cerevisiaeMutant With a Constitutively Activated cAMP-Dependent Pathway
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Genome-Wide Analysis of the Effects of Heat Shock on aSaccharomyces cerevisiaeMutant With a Constitutively Activated cAMP-Dependent Pathway

机译：全基因组分析热休克对啤酒酵母突变体具有组成性激活的cAMP依赖性途径的影响。

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We have used DNA microarray technology and 2-D gel electrophoresis combined withmass spectrometry to investigate the effects of a drastic heat shock from 30℃ to 50℃on a genome-wide scale. This experimental condition is used to differentiate betweenwild-type cells and those with a constitutively active cAMP-dependent pathway inSaccharomyces cerevisiae. Whilst more than 50% of the former survive this shock,almost all of the latter lose viability. We compared the transcriptomes of the wildtypeand a mutant strain deleted for the genePDE2, encoding the high-affinity cAMPphosphodiesterase before and after heat shock treatment. We also compared the twoheat-shocked samples with one another, allowing us to determine the changes thatoccur in thepde2Δ mutant which cause such a dramatic loss of viability after heatshock. Several genes involved in ergosterol biosynthesis and carbon source utilizationhad altered expression levels, suggesting that these processes might be potentialfactors in heat shock survival. These predictions and also the effect of the differentphases of the cell cycle were confirmed by biochemical and phenotypic analyses. 146genes of previously unknown function were identified amongst the genes with alteredexpression levels and deletion mutants in 13 of these genes were found to be highlysensitive to heat shock. Differences in response to heat shock were also observed atthe level of the proteome, with a higher level of protein degradation in the mutant, asrevealed by comparing 2-D gels of wild-type and mutant heat-shocked samples andmass spectrometry analysis of the differentially produced proteins.
机译：我们已使用DNA微阵列技术和二维凝胶电泳结合质谱技术研究了从30℃到50℃的剧烈热休克对全基因组范围的影响。该实验条件用于区分酿酒酵母中的野生型细胞和具有cAMP组成型活性途径的细胞。尽管前者有50％以上能幸免于难，但后者几乎都失去了生存能力。我们比较了野生型的转录组和缺失基因PDE2的突变菌株，该菌株编码热激处理前后的高亲和力cAMP磷酸二酯酶。我们还将两个热激样品进行了相互比较，从而使我们能够确定在pde2Δ突变体中发生的变化，这些变化会导致热激后细胞活力的急剧下降。麦角固醇的生物合成和碳源利用中涉及的几个基因改变了表达水平，表明这些过程可能是热休克生存的潜在因素。这些预测以及细胞周期不同阶段的影响已通过生化和表型分析得到证实。在表达水平发生改变的基因中鉴定出146个以前未知功能的基因，发现其中13个基因的缺失突变体对热激高度敏感。通过比较野生型和突变体热休克样品的二维凝胶以及对产生的差异产生的质谱进行分析，揭示了在蛋白质组水平上对热激反应的差异，突变体中蛋白质降解的水平更高。蛋白质。

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