PDF(Pubmed)
Abstract：
Strategies have not been available until recently to uncover interacting protein networks specific to key cell types, their subcellular compartments, and their major regulators during complex in vivo events. Here, we apply BioID2 proximity labeling to capture protein networks acting within cardiomyocytes during a key model of innate heart regeneration in zebrafish. Transgenic zebrafish expressing a promiscuous BirA2 localized to the entire myocardial cell or membrane compartment were generated, each identifying distinct proteomes in adult cardiomyocytes that became altered during regeneration. BioID2 profiling for interactors with ErbB2, a co-receptor for the cardiomyocyte mitogen Nrg1, implicated Rho A as a target of ErbB2 signaling in cardiomyocytes. Blockade of Rho A during heart regeneration, or during cardiogenic stimulation by the mitogenic influences Nrg1, Vegfaa, or vitamin D, disrupted muscle creation. Our findings reveal proximity labeling as a useful resource to interrogate cell proteomes and signaling networks during tissue regeneration in zebrafish.
摘要：
直到最近才有策略来发现特定于关键细胞类型的相互作用蛋白质网络，它们的亚细胞区室，以及它们在复杂体内事件中的主要调节因子。这里,在斑马鱼先天性心脏再生的关键模型中，我们应用BioID2邻近标记捕获作用于心肌细胞内的蛋白质网络。产生了表达位于整个心肌细胞或膜区室的混杂BirA2的转基因斑马鱼，每个鉴定在再生过程中发生改变的成年心肌细胞中不同的蛋白质组。与心肌细胞有丝分裂原Nrg1的共受体ErbB2的相互作用物的BioID2分析表明RhoA是心肌细胞中ErbB2信号传导的靶标。在心脏再生过程中阻断RhoA，或在有丝分裂影响Nrg1，Vegfaa的心源性刺激期间，或者维生素D,破坏肌肉的产生。我们的发现表明，邻近标记是斑马鱼组织再生过程中询问细胞蛋白质组和信号网络的有用资源。
