Global gene expression in recombinant and non-recombinant yeast Saccharomyces cerevisiae in three different metabolic states.
Global gene expression in recombinant and non-recombinant yeast Saccharomyces cerevisiae in three different metabolic states.
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DOI：
10.1016/j.biotechadv.2009.05.015
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发表时间：
2009-11
影响因子：
16
通讯作者：
H. Díaz;B. Andrews;A. Hayes;J. Castrillo;S. Oliver;J. Asenjo
中科院分区：
文献类型：
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作者：
H. Díaz;B. Andrews;A. Hayes;J. Castrillo;S. Oliver;J. Asenjo
两种酿酒酵母菌株的整体基因表达，一种是积累大量细胞内蛋白质超氧化物歧化酶（SOD）的重组体（P+），另一种是不含重组质粒的非重组体（P-），当细胞在葡萄糖上指数生长时，而在使用甘油时的早期稳定期。当比较在乙醇和葡萄糖（Eth/Gluc）上生长期间P−（和P+）的基因表达时，过表达与甘油消耗的增加、TCA循环的激活、糖原的降解和乙醇的代谢有关。此外，97.6%的基因（80个基因）参与的中央代谢途径过表达。这与DeRisi等人[DeRisi，J.L.，Iyer，V.R. & Brown，P.O. 1997.在基因组尺度上探索基因表达的代谢和遗传控制。科学278：680-686。但与代谢通量分析（MFA）所观察到的非常不同，其中比生长速率降低至约100%。40%，TCA循环中的通量降低到约100%。40%（在P+中为30%），糖酵解减少到几乎为0，蛋白质合成减少到约40%。50%（P+为40%）。显然，不可能以简单或直接的方式将定量mRNA表达水平与代谢通量分析（MFA）显示的细胞功能相关联。当在3个生长阶段比较两个菌株时，发现在所有情况下有4个基因表达不足或过表达。所有这些基因的产物都在酵母的质膜或细胞壁上表达。当比较在葡萄糖、乙醇和早期稳定期生长的菌株（P+/P-）时，许多中心代谢途径的基因在P+中表达不足，这与两种菌株（MFA）的代谢通量的行为相似。比较早期稳定期与乙醇生长期（Stat/Eth）的P-（和某种程度上的P+）基因表达，普遍存在表达不足。这表明，乙醇和早期稳定期之间的代谢转换对基因表达具有几乎瞬时的影响，但对代谢通量具有更延迟的影响，并且从代谢分析的角度来看，“早期稳定”期代表“晚期乙醇”期，因为乙醇仍然存在并被消耗，尽管速度慢得多。
Global gene expression of two strains of Saccharomyces cerevisiae, one recombinant (P+), accumulating large amounts of an intracellular protein Superoxide Dismutase (SOD) and one non-recombinant (P−) which does not contain the recombinant plasmid, were compared in batch culture during diauxic growth when cells were growing exponentially on glucose, when they were growing exponentially on ethanol, and in the early stationary phase when glycerol was being utilized. When comparing the gene expression for P− (and P+) during growth on ethanol to that on glucose (Eth/Gluc), overexpression is related to an increase in consumption of glycerol, activation of the TCA cycle, degradation of glycogen and metabolism of ethanol. Furthermore, 97.6% of genes (80 genes) involved in the central metabolic pathway are overexpressed. This is similar to that observed by DeRisi et al. [DeRisi, J.L., Iyer, V.R. & Brown, P.O. 1997. Exploring the metabolic and genetic control of gene expression on a genomic scale. Science 278:680–686.] but very different from was observed for Metabolic Flux Analysis (MFA), where the specific growth rate is lowered to ca. 40%, the fluxes in the TCA cycle are reduced to ca. 40% (to 30% in P+), glycolysis is reduced to virtually 0 and protein synthesis to ca. 50% (to 40% in P+). Clearly it is not possible to correlate in a simple or direct way, quantitative mRNA expression levels with cell function which is shown by the Metabolic Flux Analysis (MFA). When comparing the two strains in the 3 growth stages, 4 genes were found to be under or overexpressed in all cases. The products of all of these genes are expressed at the plasma membrane or cell wall of the yeast. While comparing the strains (P+/P−) when growing on glucose, ethanol and in the early stationary phase, many of the genes of the central metabolic pathways are underexpressed in P+, which is similar to the behaviour of the metabolic fluxes of both strains (MFA). Comparing the gene expression for P− (and to some extent P+) during the early stationary phase to growth on ethanol (Stat/Eth), underexpression is generalized. This shows that the switch in metabolism between ethanol and early stationary phases has an almost instantaneous effect on gene expression but a much more retarded effect on metabolic fluxes and that the “early stationary” phase represents a “late ethanol” phase from the metabolic analysis point of view since ethanol is still present and being consumed although at a much slower rate.