Transcriptional profiling of Saccharomyces cerevisiae exposed to propolis.
Transcriptional profiling of Saccharomyces cerevisiae exposed to propolis.
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DOI：
10.1186/1472-6882-12-194
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发表时间：
2012-10-24
影响因子：
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通讯作者：
Goldman GH
中科院分区：
文献类型：
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作者：
de Castro PA;Savoldi M;Bonatto D;Malavazi I;Goldman MH;Berretta AA;Goldman GH
蜂胶是蜜蜂（Apis mellifera）从各种植物来源收集的植物树脂的天然产物。我们以前的研究表明，酵母细胞对线粒体的敏感性依赖于线粒体的功能，空泡酸化和自噬在酵母细胞死亡中起重要作用。在这里，我们扩展了我们的理解蜂胶介导的细胞死亡的酵母酿酒酵母，通过应用系统生物学工具来分析暴露于蜂胶的细胞的转录谱。我们使用了S.酿酒酵母菌暴露于乙醇中。我们通过使用选定基因的实时PCR验证了我们的发现。应用系统生物学工具（物理蛋白质-蛋白质相互作用[PPPI]网络）分析蜂胶诱导的转录活性，旨在确定哪些途径受蜂胶S.酿酒酵母和潜在地影响细胞死亡。当与参考时间（未处理的样品）相比时，我们能够观察到在至少一个时间点中调节的1，339个基因（t检验，p值0.01）。通过基因本体（GO）术语查找器工具进行的富集分析显示在微阵列杂交中上调的基因中的几个生物学类别的富集，例如转运和跨膜转运以及对应激的响应。实时荧光定量PCR分析表明，我们的微阵列杂交方法所选择的基因能够提供有关S。酿酒酵母基因表达调控具有相当高的置信水平。最后，物理蛋白质-蛋白质（PPPI）网络设计和全局拓扑分析强调了这些途径在S。酿酒酵母的转录数据与通过微阵列分析获得的转录数据相关联。总之，我们的数据表明，在很大程度上影响真核细胞中的几个途径。然而，最突出的途径与氧化应激、线粒体电子传递链、液泡酸化、与蛋白质靶向液泡相关的大自噬调节、细胞对饥饿的反应以及RNA聚合酶II启动子转录的负调节有关。我们的工作再次强调了S.酿酒酵母作为模型系统，以在分子水平上理解在本文测试的浓度下，酵母菌引起该生物体中细胞死亡的机制。我们的研究是第一个通过使用功能基因组学系统地研究蜂胶如何影响和调节生物体的mRNA丰度，并可能刺激进一步研究真菌中蜂胶介导的细胞死亡机制的研究。
Propolis is a natural product of plant resins collected by honeybees (Apis mellifera) from various plant sources. Our previous studies indicated that propolis sensitivity is dependent on the mitochondrial function and that vacuolar acidification and autophagy are important for yeast cell death caused by propolis. Here, we extended our understanding of propolis-mediated cell death in the yeast Saccharomyces cerevisiae by applying systems biology tools to analyze the transcriptional profiling of cells exposed to propolis. We have used transcriptional profiling of S. cerevisiae exposed to propolis. We validated our findings by using real-time PCR of selected genes. Systems biology tools (physical protein-protein interaction [PPPI] network) were applied to analyse the propolis-induced transcriptional bevavior, aiming to identify which pathways are modulated by propolis in S. cerevisiae and potentially influencing cell death. We were able to observe 1,339 genes modulated in at least one time point when compared to the reference time (propolis untreated samples) (t-test, p-value 0.01). Enrichment analysis performed by Gene Ontology (GO) Term finder tool showed enrichment for several biological categories among the genes up-regulated in the microarray hybridization such as transport and transmembrane transport and response to stress. Real-time RT-PCR analysis of selected genes showed by our microarray hybridization approach was capable of providing information about S. cerevisiae gene expression modulation with a considerably high level of confidence. Finally, a physical protein-protein (PPPI) network design and global topological analysis stressed the importance of these pathways in response of S. cerevisiae to propolis and were correlated with the transcriptional data obtained thorough the microarray analysis. In summary, our data indicate that propolis is largely affecting several pathways in the eukaryotic cell. However, the most prominent pathways are related to oxidative stress, mitochondrial electron transport chain, vacuolar acidification, regulation of macroautophagy associated with protein target to vacuole, cellular response to starvation, and negative regulation of transcription from RNA polymerase II promoter. Our work emphasizes again the importance of S. cerevisiae as a model system to understand at molecular level the mechanism whereby propolis causes cell death in this organism at the concentration herein tested. Our study is the first one that investigates systematically by using functional genomics how propolis influences and modulates the mRNA abundance of an organism and may stimulate further work on the propolis-mediated cell death mechanisms in fungi.