基于高通量微流技术的酵母细胞蛋白动力学图谱及调控网络研究

Cells response to extracellular signals through intercellular signal pathways. Those responses such as cell stress response, chemotaxis and mating response involve hundreds of proteins as the protein regulation network to realize the cell functions. To help the cell adapt to the new environments, the reactions of the proteins involved in the signaling pathway often have dynamic feature. Systematic studies of protein dynamic expression pattern after external stimulate may confine the possible signaling pathway networks and reveal the underlined mechanisms about the signal networks. In order to studies the signaling pathway such as stress responses and mating response of budding yeast, we plan to design a novel microfluidic system. Combined with time lapse microscope, we will try to realize high-throughput protein dynamic expression study of the whole signaling pathway. By this method, we can get the protein dynamic expression patterns of different signaling pathway such as stress responses and mating responses to help us to understand the biologic processes. By classified the dynamic patterns and using minimum network constraints, we may understand the control network structure and functions of the signaling pathways. These studies should also help us to understand the control mechanical of biologic system responses to the environment changes.

细胞往往通过内部的蛋白调控网络来应对外界的环境变化。这些复杂的细胞响应行为例如应激响应,化学趋向性,细胞杂交等,往往涉及上百个蛋白作为蛋白调控网络来实现相应的生物功能。这些蛋白的响应以一定的蛋白时间序列的变化来体现,使得细胞能更好的适应外界环境的变化。系统的研究加入刺激后的蛋白动力学图谱可以限制可能的信号通路甚至揭示背后的调控网络机制。我们计划以酵母细胞应激响应通路以及mating响应通路网络研究为例，设计新型的高通量的微流系统，通过时序显微镜并行记录多维度的芽殖酵母细胞表型。通过这些数据，我们可以获得不同的生物功能下游蛋白的动力学图谱，对其生物学轨迹进行聚类。结合已知的一些生物学知识，以及最小网络的数学办法，我们可能可以解析这些信号通路下游的蛋白相互作用网络结构以及功能的关系，也将为理解生物系统对外界环境刺激的响应机制提供较大的帮助。

细胞往往通过内部的蛋白调控网络来应对外界的环境变化。这些复杂的细胞响应行为例如应激响应,化学趋向性,细胞杂交等,往往涉及几十上百个蛋白作为蛋白调控网络来实现相应的生物功能。这些蛋白的响应以一定的蛋白时间序列的变化来体现,使得细胞能更好的适应外界环境的变化。我们通过本项目，对酵母细胞交配应激通路，渗透压响应型号通路，孢子通路以及衰老过程中的应激能力变化进行了系统性的研究，完成了项目原先预定目标，发表包括PNAS, PRL, Plos computational Biology, Lab on a chip, Advanced Materials Technologies， Scientific reports, Integrative Biology等SCI论文14篇，申请两项国家发明专利。我们的工作极大的推动了相关定量生物学的研究进展。