Pellino-2 is an E3 ubiquitin ligase that mediates intracellular signaling in innate immune pathways. Most studies of endogenous Pellino-2 have been performed in macrophages, but none in nonimmune cells. Using yeast two-hybrid screening and co-immunoprecipitation, we identified six novel interaction partners of Pellino-2, with various localizations: insulin receptor substrate 1, NIMA-related kinase 9, tumor necrosis factor receptor-associated factor 7, cyclin-F, roundabout homolog 1, and disheveled homolog 2. Pellino-2 showed cytoplasmic localization in a wide range of nonimmune cells under physiological potassium concentrations. Treatment with the potassium ionophore nigericin resulted in nuclear localization of Pellino-2, which was reversed by the potassium channel blocker tetraethylammonium. Live-cell imaging revealed intracellular migration of GFP-tagged Pellino-2. In summary, Pellino-2 interacts with proteins at different cellular locations, taking part in dynamic processes that change its intracellular localization influenced by potassium efflux.

中文翻译：

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非免疫细胞中的 Pellino-2：新的相互作用伙伴和细胞内定位

Pellino-2 是一种 E3 泛素连接酶，可介导先天免疫途径中的细胞内信号传导。大多数内源性 Pellino-2 的研究都是在巨噬细胞中进行的，但没有在非免疫细胞中进行。使用酵母双杂交筛选和免疫共沉淀，我们确定了 Pellino-2 的六个新的相互作用伙伴，具有不同的定位：胰岛素受体底物 1、NIMA 相关激酶 9、肿瘤坏死因子受体相关因子 7、细胞周期蛋白-F、迂回的同源物 1 和蓬乱的同源物 2。Pellino-2 在生理钾浓度下在广泛的非免疫细胞中显示出细胞质定位。用钾离子载体尼日利亚菌素处理导致 Pellino-2 的核定位，这被钾通道阻滞剂四乙铵逆转。活细胞成像揭示了 GFP 标记的 Pellino-2 的细胞内迁移。总之，Pellino-2 与不同细胞位置的蛋白质相互作用，参与改变其受钾外流影响的细胞内定位的动态过程。