Upon activation by different transmembrane receptors, the same signaling protein can induce distinct cellular responses. A way to decipher the mechanisms of such pleiotropic signaling activity is to directly manipulate the decision-making activity that supports the selection between distinct cellular responses. We developed an optogenetic probe (optoSRC) to control SRC signaling, an example of a pleiotropic signaling node, and we demonstrated its ability to generate different acto-adhesive structures (lamellipodia or invadosomes) upon distinct spatio-temporal control of SRC kinase activity. The occurrence of each acto-adhesive structure was simply dictated by the dynamics of optoSRC nanoclusters in adhesive sites, which were dependent on the SH3 and Unique domains of the protein. The different decision-making events regulated by optoSRC dynamics induced distinct downstream signaling pathways, which we characterized using time-resolved proteomic and network analyses. Collectively, by manipulating the molecular mobility of SRC kinase activity, these experiments reveal the pleiotropy-encoding mechanism of SRC signaling.

在被不同的跨膜受体激活后，相同的信号蛋白可以诱导不同的细胞反应。破译这种多效性信号活动机制的一种方法是直接操纵支持不同细胞反应之间选择的决策活动。我们开发了一种光遗传学探针 (optoSRC) 来控制 SRC 信号传导，这是多效性信号传导节点的一个例子，并且我们证明了它能够在对 SRC 激酶活性进行不同时空控制时产生不同的行为粘附结构（片状伪足或侵袭体）。每个acto-adhesive结构的出现简单地由粘附位点中的optoSRC纳米团簇的动力学决定，这取决于蛋白质的SH3和Unique结构域。由 optoSRC 动力学调节的不同决策事件诱导了不同的下游信号通路，我们使用时间分辨蛋白质组学和网络分析对其进行了表征。总的来说，通过操纵 SRC 激酶活性的分子迁移率，这些实验揭示了 SRC 信号传导的多效性编码机制。