Amezquita Jonathan, Desbois Muriel, Opperman Karla J, Pak Joseph S, Christensen Elyse L, Nguyen Nikki T, Diaz-Garcia Karen, Borgen Melissa A, Grill Brock
Center for Integrative Brain Research, Seattle Children's Research Institute, Seattle, WA, USA.
Molecular and Cellular Biology Graduate Program, University of Washington, Seattle, WA, USA.
bioRxiv. 2023 Nov 15:2023.11.15.566604. doi: 10.1101/2023.11.15.566604.
Integrin signaling plays important roles in development and disease. An adhesion signaling network called the integrin adhesome has been principally defined using bioinformatics and proteomics. To date, the adhesome has not been studied using integrated proteomic and genetic approaches. Here, proteomic studies in identified physical associations between the RPM-1 ubiquitin ligase signaling hub and numerous adhesome components including Talin, Kindlin and beta-integrin. RPM-1 is orthologous to human MYCBP2, a prominent player in nervous system development associated with a neurodevelopmental disorder. Using neuron-specific, CRISPR loss-of-function strategies, we show that core adhesome components affect axon development and interact genetically with RPM-1. Mechanistically, Talin opposes RPM-1 in a functional 'tug-of-war' on growth cones that is required for accurate axon termination. Thus, our findings orthogonally validate the adhesome via multi-component genetic and physical interfaces with a key neuronal signaling hub and identify new links between the adhesome and brain disorders.
整合素信号传导在发育和疾病中发挥着重要作用。一个名为整合素粘附体的粘附信号网络主要是通过生物信息学和蛋白质组学来定义的。迄今为止，尚未使用综合蛋白质组学和遗传学方法对粘附体进行研究。在这里，研究中的蛋白质组学研究确定了RPM-1泛素连接酶信号枢纽与许多粘附体成分之间的物理关联，这些成分包括踝蛋白、桩蛋白和β-整合素。RPM-1与人类MYCBP2直系同源，MYCBP2是神经系统发育中的一个重要参与者，与一种神经发育障碍有关。使用神经元特异性的CRISPR功能丧失策略，我们表明核心粘附体成分影响轴突发育，并与RPM-1发生遗传相互作用。从机制上讲，踝蛋白在生长锥上与RPM-1进行功能性的“拔河比赛”，这是准确轴突终止所必需的。因此，我们的研究结果通过与一个关键神经元信号枢纽的多组分遗传和物理界面正交验证了粘附体，并确定了粘附体与脑部疾病之间的新联系。
