Microtubule-dependent ribosome localization in C. elegans neurons.
Microtubule-dependent ribosome localization in C. elegans neurons.
复制标题
作者：
Noma K;Goncharov A;Ellisman MH;Jin Y
核糖体的亚细胞定位定义了蛋白质合成的位置和能力。在核糖体动力学的细胞生物学机制研究中，需要体内可视化多细胞生物中核糖体的方法。在这里，我们开发了一种使用 split GFP 进行秀丽隐杆线虫核糖体组织特异性可视化的方法。标记的核糖体在特定神经元类型的轴突和突触末端中被检测为荧光点，与超微结构水平的核糖体分布相关。我们发现轴突核糖体在神经元发育过程中和轴突损伤后改变定位。通过检查影响轴突运输的突变体并进行正向遗传筛选，我们发现微管细胞骨架和 JIP3 蛋白 UNC-16 对轴突和体细胞核糖体的定位具有明显的影响。我们的数据证明了体内核糖体组织特异性可视化的实用性，并提供了对神经元中核糖体定位主动调节机制的深入了解。
Subcellular localization of ribosomes defines the location and capacity for protein synthesis. Methods for in vivo visualizing ribosomes in multicellular organisms are desirable in mechanistic investigations of the cell biology of ribosome dynamics. Here, we developed an approach using split GFP for tissue-specific visualization of ribosomes in Caenorhabditis elegans. Labeled ribosomes are detected as fluorescent puncta in the axons and synaptic terminals of specific neuron types, correlating with ribosome distribution at the ultrastructural level. We found that axonal ribosomes change localization during neuronal development and after axonal injury. By examining mutants affecting axonal trafficking and performing a forward genetic screen, we showed that the microtubule cytoskeleton and the JIP3 protein UNC-16 exert distinct effects on localization of axonal and somatic ribosomes. Our data demonstrate the utility of tissue-specific visualization of ribosomes in vivo, and provide insight into the mechanisms of active regulation of ribosome localization in neurons.