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LSm4 Associates with the Plasma Membrane and Acts as a Co-factor in Cell Volume Regulation

机译:LSm4与血浆膜结合,并在细胞体积调节中充当辅助因子

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ICIn is a ubiquitous, multifunctional protein with functions in cell volume regulation and RNA processing, and is thus part of an intricate protein network critically involved in the homoeostasis of cells. To better understand this vital protein network in health and disease it is fundamental to characterize the interactions between the physiological pathways in which ICln is involved, as well as the spatio-temporal regulation of these interactions. In this study, we focused on the interaction between the two best studied pathways in which ICln is involved - regulatory volume decrease and RNA processing - and asked, whether or not the RNA processing factor and ICln interaction partner LSm4 may also have a function in cell volume regulation in NIH3T3 fibroblasts or HEK293 Phoenix cells. To address this question, we studied in isotonic and hypotonic conditions by FRET, biochemistry and electrophysiology, the intracellular distribution of the RNA processing factor LSm4, its interaction with ICln, as well as the involvement of LSm4 in the activation of the swelling dependent anion and osmolyte channel ICIswell. In isotonic conditions, LSm4 associates with ICln, and the plasma membrane. Hypotonic cell swelling leads to the dissociation of LSm4 from the plasma membrane, and from ICln. Over-expression of LSm4 affects the translocation of ICln to the cell membrane and markedly inhibits the activation kinetics and current density of ICIswell. These findings indicate that LSm4 not only acts in RNA processing, but also as a co-factor in cell volume regulation.
机译:ICIn是一种普遍存在的多功能蛋白,在细胞体积调节和RNA加工中具有功能,因此是复杂蛋白质网络的一部分,该网络至关重要地参与了细胞的同质化。为了更好地理解健康和疾病中的这一至关重要的蛋白质网络,表征参与ICln的生理途径之间的相互作用以及这些相互作用的时空调节至关重要。在这项研究中,我们着重研究了涉及ICln的两个最佳研究途径之间的相互作用-调节量减少和RNA加工-并询问RNA加工因子和ICln相互作用伴侣LSm4是否在细胞中也有功能NIH3T3成纤维细胞或HEK293 Phoenix细胞中的血容量调节。为了解决这个问题,我们在等渗和低渗条件下,通过FRET,生物化学和电生理学,RNA加工因子LSm4的细胞内分布,其与ICln的相互作用以及LSm4参与溶胀性负离子和水分子的活化研究。渗透液通道ICIswell。在等渗条件下,LSm4与ICln和质膜结合。低渗性细胞肿胀导致LSm4从质膜和ICln解离。 LSm4的过表达影响ICln到细胞膜的转运,并显着抑制ICIswell的激活动力学和电流密度。这些发现表明,LSm4不仅在RNA加工中起作用,而且还作为细胞体积调节的辅助因子。

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