Optogenetic Dissection of Protein Kinase Networks

细胞对外界刺激的反应是由许多信号通路控制的，这些信号通路本身相互连接形成信号网络。激酶是进行位点特异性磷酸化的酶，这是信号网络中信息传递的主要形式。SPARKLE是一种利用光来控制激酶活性的方法;基因编码的激酶被改造成在其活性位点含有笼状赖氨酸残基，并且这些酶只有在暴露于紫外线后才变得活跃。通过使用光来实现特定激酶激活的空间和时间特异性，可以解剖由这些激酶控制的细胞子网络的活性。通过这种方法，将有可能阐明信号通路之间的串扰和反馈机制，并定义特定通路在驱动细胞极化和定向运动中的作用。更广泛的影响：这个项目将提供化学，细胞生物学和计算建模的本科生和研究生的跨学科培训。一个新的实验室为基础的光遗传学课程将被开发，光在生物系统中的作用将在罗利的大理石儿童博物馆进行推广。该奖项由分子和细胞生物科学部的细胞动力学和功能集群以及化学部的生命过程化学计划共同支持。

Cell responses to external stimuli are controlled by numerous signaling pathways that are themselves interconnected to form signaling networks. Kinases are enzymes that carry out site-specific phosphorylation, the primary form of information transfer in signaling networks. SPARKLE is an approach to controlling the activity of kinases using light; genetically-encoded kinases are engineered to contain a caged lysine residue at their active site, and these enzymes then become active only after exposure to ultraviolet light. By using light to achieve spatial and temporal specificity in the activation of particular kinases, the activity of the cellular sub-networks controlled by these kinases can be dissected. With this approach it will be possible to elucidate cross-talk and feedback mechanisms between signaling pathways, and to define the role of particular pathways in driving the polarization and directed movement of cells. Broader Impacts: This project will provide interdisciplinary training in chemistry, cell biology, and computational modeling to undergraduate and graduate students. A new laboratory-based course in optogenetics will be developed, and outreach on the role of light in biological systems will be conducted at the Marbles Kids Museum in Raleigh.This award is supported jointly by the Cellular Dynamics and Function Cluster of the Molecular and Cellular Biosciences Division, and by the Chemistry of Life Processes Program of the Division of Chemistry.