Elucidating the interplay of PMF determinants in mitochondria and chloroplasts in folio

线粒体和叶绿体一直在塑造我们所知的真核生物，质子动力（PMF）是这两种细胞器功能的核心。PMF将电子传递的氧化还原化学与ATP合成联系起来，但也作为一个调节中心，决定了细胞器的功能和组成。在植物和藻类中，线粒体和叶绿体的PMF通过细胞代谢网络相互连接。虽然有效的光合作用需要叶绿体和线粒体生物能量学协同工作，但这种相互作用在体内是如何实现和支撑的仍然没有得到充分的理解。在P07中，我们将阐明质子（H+），钾离子（K+）和钙离子（Ca 2+）的亚细胞动力学如何决定体内线粒体和叶绿体中的PMF特性。以拟南芥成熟莲座叶为模型，我们将利用一套基于荧光蛋白的生物传感器和高分辨率成像技术来生成拟南芥叶绿体和线粒体亚室中H+、K+和Ca2+的图谱。该图谱不仅包括活叶组织的三维分子信息，还包括对不同光照条件的响应。它将提供一个迄今为止唯一的参考，并将被用来进一步证实候选蛋白的H+，K+和Ca2+跨细胞器膜运输的意义，在确定PMF的特点，在pneumonia。选择的线粒体和叶绿体的候选蛋白质的遗传突变体，以设置细胞器H+，K+和Ca2+的动态将测试其对细胞器PMF特性的影响，在ESTA。其中几个参与者的作用，如KEA3和NHD1，将在该联盟内密切合作进行评估，而其他参与者的作用，如MCU，MICU，UCP1和AOX1a将根据我们实验室的最新进展进行评估。图谱的方法和突变体相结合，将允许解决的核心问题，在何种程度上在一个细胞器中的PMF特性的变化影响其他。改变PMF动态的光合代谢，细胞器超微结构和整个植物的性能的意义将通过与联盟内的领先专家的合作变得可访问。作为结果，将建立对体内PMF调节和特定候选蛋白质参与者在设定PMF特征中的相对重要性的先进理解。

Mitochondria and chloroplasts have been shaping eukaryotic life as we know it. At the center of the function of both organelles is the proton motive force (PMF). The PMF connects redox chemistry of electron transport with ATP synthesis, but also acts as a regulation hub that determines the function and the makeup of the organelle. In plants and algae, the PMF of both mitochondria and chloroplasts are interconnected via the cellular metabolic network. While efficient photosynthesis requires chloroplast and the mitochondrial bioenergetics to work in concert, how exactly this interplay is achieved and underpinned in vivo remains insufficiently understood. In P07, we will elucidate how the subcellular dynamics of protons (H+), potassium ions (K+) and calcium ions (Ca2+) determine PMF characteristics in mitochondria and chloroplasts in vivo. Using a mature rosette leaf of Arabidopsis thaliana as a model, we will exploit a set of fluorescent protein-based biosensors and high-resolution imaging to generate an atlas of H+, K+ and Ca2+ in the subcompartments of the chloroplasts and the mitochondria in folio. The atlas will not only comprise three-dimensional molecular information in living leaf tissues, but also responses to different illumination regimes. It will provide a hitherto unique reference and will be used to further corroborate the significance of candidate proteins for H+, K+ and Ca2+ transport across the organelle membranes in determining PMF characteristics in folio. A selection of genetic mutants of candidate proteins of the mitochondria and the chloroplasts to set organelle H+, K+ and Ca2+ dynamics will be tested for their impact on organellar PMF characteristics in folio. The role of several of those players, such as KEA3 and NHD1, will be assessed in close collaboration within this consortium, while that of others such as the MCUs, MICU, UCP1 and AOX1a will be assessed based on the recent advances by our lab. The atlas approach and the mutants combined will allow to address the central question to what extent changes in the PMF characteristics in one organelle affect the other. The significance of altered PMF dynamics for photosynthetic metabolism, organelle ultrastructure, and whole plant performance will become accessible through collaboration with leading experts within the consortium. An advanced understanding of PMF regulation in vivo and the relative significance of specific candidate protein players in setting PMF characteristics will be established as a result.