SFB 1381: Dynamic organization of cellular protein machineries: From biogenesis and modular assembly to function

复杂的生物功能是由充当分子机器的多蛋白组件执行的。它们的功能涵盖了细胞生理的方方面面，从DNA的复制、修复和转录，到蛋白质的翻译、折叠和降解，能量的转换和分子的运输，以及细胞器和细胞之间通过信号转导进行的通讯。虽然技术进步导致识别出过多的不同亚基和蛋白质-蛋白质相互作用，但挑战仍然是识别单个亚基逐步组装成大分子复合体，以及这些组装提供的功能收益远远超过它们各自蛋白质的总和。此外，蛋白质机械不是静态的实体，而是可以动态地与其他复合体结合形成组装网络。它们也可以部分或完全分解，以响应配体或第二信使的结合或细胞代谢状态的变化。在CRC1381中，我们的目标是解决蛋白质机械动态组织的挑战性开放问题。我们的跨学科方法结合了分子和细胞生物学、生物物理学和结构生物学，使我们能够在分子水平上全面分析复杂蛋白质机器的生物发生和组装，直到研究细胞需求对它们施加的功能输出和调控控制。凭借这一独特的专业知识，CRC1381研究了i)多蛋白质机器在生物发生过程中如何组装成功能单元，ii)模块化组件如何影响功能，以及iii)这些组件如何集成到动态细胞网络中并受外部刺激控制以使细胞适应细胞需求。在第一个资助期，我们对被调查的机制的组成产生了详细的分子洞察力：我们发现了缺失的蛋白质成分，并确定了调节不同功能输出的新型亚组分。在第二个资助期，我们希望将我们的分子和机制分析扩展到细胞动力学和细胞信号的整合，以研究在空间和时间细胞背景下组装的蛋白质机制及其不同的模块。为此，我们将把我们的核心专业知识与先进的显微技术相结合，以解决如何在细胞内不同位置动态形成子模块、如何通过信号通路调节组装和拆卸以及细胞蛋白质机制如何使其功能适应细胞状态变化的问题。在我们多学科专业知识的基础上，我们提出了一种全面的、高度整合的研究方法，以揭示控制生命细胞过程的多蛋白组件的生物发生和功能机制。

Complex biological functions are performed by multiprotein assemblies that act as molecular machineries. Their functions encompass all aspects of cellular physiology from DNA replication, repair and transcription to protein translation, folding and degradation, the conversion of energy and the transport of molecules, to the communication via signal transduction between organelles and cells. While technological advancements resulted in the identification of a plethora of different subunits and protein-protein interactions, the challenge remains to identify the step-wise assembly of the individual subunits into macromolecular complexes and the functional gain that these assemblies provide that is far more than the sum of their individual proteins. In addition, protein machineries are not static entities, but can dynamically associate with other complexes to form a network of assemblies. They can also partially or fully disassemble in response to binding of ligands or second messengers or changes within the metabolic state of the cell. In the CRC1381 we aim to address the challenging open questions of the dynamic organization of protein machineries. Our interdisciplinary approach combines molecular and cell biology, biophysics, and structural biology and enables us to comprehensively analyze the biogenesis and assembly of complex protein machineries on the molecular level up to the investigation of the functional outputs and regulatory controls that cellular demands exert on them. With this unique expertise the CRC1381 investigates i) how multiprotein machineries are assembled into functional units during biogene-sis, ii) how modular assemblies impact on function and iii) how these assemblies are integrated into dynamic cellular networks and controlled by external stimuli to adapt the cell to cellular demands. In the first funding period we generated detailed molecular insights into the composition of the investigated machineries: We uncovered missing protein components and identified novel subassemblies that mediate different functional outputs. In the second funding period we want to expand our molecular and mechanistic analysis towards integration of cellular dynamics and cellular signaling to investigate the assembled protein machineries and their different modules in the spatial and temporal cellular context. For this we will combine our core expertise with advanced microscopy techniques to address the questions how submodules are formed dynamically and at different localizations in the cell, how assembly and disassembly is regulated by signaling pathways, and how cellular protein machineries adapt their function to changes in cellular states. Building on our multidisciplinary expertise, we propose a comprehensive, and highly integrative research approach to uncover the mechanisms in biogenesis and function of multiprotein assemblies that govern the cellular processes of life.