Snijders Ambrosius P L, Walther Jasper, Peter Stefan, Kinnman Iris, de Vos Marjon G J, van de Werken Harmen J G, Brouns Stan J J, van der Oost John, Wright Phillip C
Biological and Environmental Systems Group, Department of Chemical and Process Engineering, University of Sheffield, Sheffield, UK.
Proteomics. 2006 Mar;6(5):1518-29. doi: 10.1002/pmic.200402070.
In the last decade, an increasing number of sequenced archaeal genomes have become available, opening up the possibility for functional genomic analyses. Here, we reconstructed the central carbon metabolism in the hyperthermophilic crenarchaeon Sulfolobus solfataricus (glycolysis, gluconeogenesis and tricarboxylic acid cycle) on the basis of genomic, proteomic, transcriptomic and biochemical data. A 2-DE reference map of S. solfataricus grown on glucose, consisting of 325 unique ORFs in 255 protein spots, was created to facilitate this study. The map was then used for a differential expression study based on (15)N metabolic labelling (yeast extract + tryptone-grown cells (YT) vs. glucose-grown cells (G)). In addition, the expression ratio of the genes involved in carbon metabolism was studied using DNA microarrays. Surprisingly, only 3 and 14% of the genes and proteins, respectively, involved in central carbon metabolism showed a greater than two-fold change in expression level. All results are discussed in the light of the current understanding of central carbon metabolism in S. solfataricus and will help to obtain a system-wide understanding of this organism.
在过去十年中，越来越多已测序的古菌基因组可供使用，为功能基因组分析开辟了可能性。在此，我们基于基因组、蛋白质组、转录组和生化数据，重建了嗜热泉古菌嗜热栖热菌（糖酵解、糖异生和三羧酸循环）的中心碳代谢。构建了一张在葡萄糖上生长的嗜热栖热菌的二维参考图谱，该图谱由255个蛋白点中的325个独特开放阅读框组成，以促进这项研究。然后该图谱被用于基于¹⁵N代谢标记的差异表达研究（酵母提取物 + 胰蛋白胨培养的细胞（YT）与葡萄糖培养的细胞（G））。此外，使用DNA微阵列研究了参与碳代谢的基因的表达比率。令人惊讶的是，参与中心碳代谢的基因和蛋白质中，分别只有3%和14%的表达水平变化超过两倍。所有结果都根据目前对嗜热栖热菌中心碳代谢的理解进行了讨论，这将有助于从全系统角度了解这种生物。
