Subcellular analysis of metabolic network dynamics under elevated CO2

高等植物的光合作用和初级代谢受到严格的调节。当环境发生变化时，植物需要立即调整其新陈代谢，以防止慢性组织损伤和细胞死亡。植物细胞的高度区室化使环境变化中代谢调节的研究变得复杂。基于以前的研究结果，我们的项目旨在量化CO2升高（eCO2）下亚细胞代谢网络的动态，应用亚细胞分馏，组学分析和数学建模的组合。此外，该项目的目的是定量评估的作用，三种酶的差异亚细胞定位，以评估他们的作用，在这种生长条件下。以前，我们发现的证据表明，己激酶1（HXK1）在光呼吸中的作用，并与羟丙酮酸还原酶1（HPR 1）和线粒体转运蛋白A BOUT DE SOUFFLE（BOU）的潜在的调节相互作用。因此，在我们的项目中，我们的目标是破译HXK1在eCO2下初级代谢的亚细胞动力学中的作用，以揭示HXK1缺乏如何影响光呼吸途径。我们将联合收割机亚细胞代谢组学与蛋白质组学和转录组学相结合，以确定hpr1和bou突变的亚细胞后果，它们在eCO2下的相关性以及与HXK1可能的相互作用。最后，我们将开发和应用一个基因组规模的数据集成平台，用于亚细胞代谢的分析。我们将结合联合收割机转录组和蛋白质组动力学的上下文特定的网络重建和亚细胞代谢动力学的分析。这将揭示从转录组学和蛋白质组学数据重建的网络之间的相似性和差异，并将显示光呼吸代谢如何与HXK1相互作用并依赖于HXK1。

Photosynthesis and primary metabolism of higher plants are tightly regulated. Upon environmental changes, plants need to immediately adjust their metabolism in order to prevent chronic tissue damage and cell death. The study of metabolic regulation in changing environments is complicated by the high degree of compartmentalization of plant cells. Based on previous findings, our project aims to quantify dynamics of subcellular metabolic networks under elevated CO2 (eCO2), applying a combination of subcellular fractionation, omics analysis and mathematical modeling. Further, the objective of this project is to quantitatively assess the role of three enzymes with differential subcellular localization to assess their role under such growth conditions. Previously, we found evidence for a role of HEXOKINASE 1 (HXK1) in photorespiration and a potential regulatory interaction with HYDROXYPYRUVATE REDUCTASE 1 (HPR1) and the mitochondrial transporter A BOUT DE SOUFFLE (BOU). Thus, in our project, we aim to decipher the role of HXK1 for subcellular kinetics of primary metabolism under eCO2 to reveal how HXK1-deficiency affects photorespiratory pathways. We will combine subcellular metabolomics with proteomics and transcriptomics to identify subcellular consequences of the hpr1 and bou mutations, their relevance under eCO2 and possible interactions with HXK1. Finally, we will develop and apply a genome-scale data integration platform for analysis of subcellular metabolism. We will combine transcriptome and proteome dynamics for context-specific network reconstruction and for the analysis of subcellular metabolite dynamics. This will reveal similarities and discrepancies between networks reconstructed from transcriptomics and proteomics data and it will show how photorespiratory metabolism interacts with and depends on HXK1.