Institute of Molecular Systems Biology, Eidgenössische Technische Hochschule (ETH) Zürich, 8093 Zürich, Switzerland.
Institute of Molecular Life Sciences and Swiss Institute of Bioinformatics, University of Zürich, 8057 Zürich, Switzerland.
Sci Signal. 2014 Nov 25;7(353):rs6. doi: 10.1126/scisignal.2005602.
Protein phosphorylation is a widespread posttranslational modification that regulates almost all cellular functions. To investigate the large number of phosphorylation events with unknown functions, we monitored the concentrations of several hundred intracellular metabolites in Saccharomyces cerevisiae yeast strains with deletions of 118 kinases or phosphatases. Whereas most deletion strains had no detectable difference in growth compared to wild-type yeast, two-thirds of deletion strains had alterations in metabolic profiles. For about half of the kinases and phosphatases encoded by the deleted genes, we inferred specific regulatory roles on the basis of knowledge about the affected metabolic pathways. We demonstrated that the phosphatase Ppq1 was required for metal homeostasis. Combining metabolomic data with published phosphoproteomic data in a stoichiometric model enabled us to predict functions for phosphorylation in the regulation of 47 enzymes. Overall, we provided insights and testable predictions covering greater than twice the number of known phosphorylated enzymes in yeast, suggesting extensive phosphorylation-dependent regulation of yeast metabolism.
蛋白质磷酸化是一种广泛存在的翻译后修饰，调节着几乎所有的细胞功能。为了研究大量具有未知功能的磷酸化事件，我们监测了酿酒酵母中数百种细胞内代谢物的浓度，这些酵母菌株缺失了 118 种激酶或磷酸酶。尽管与野生型酵母相比，大多数缺失菌株的生长没有明显差异，但三分之二的缺失菌株的代谢谱发生了改变。对于删除基因编码的大约一半的激酶和磷酸酶，我们根据受影响的代谢途径的知识推断出了特定的调节作用。我们证明了磷酸酶 Ppq1 是金属内稳态所必需的。通过将代谢组学数据与已发表的磷酸蛋白质组学数据结合在一个计量模型中，我们能够预测 47 种酶的磷酸化调节功能。总的来说，我们提供了深入的见解和可测试的预测，涵盖了酵母中已知磷酸化酶的两倍以上，这表明了酵母代谢的广泛的磷酸化依赖性调节。
