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首页> 外文期刊>Cellular Signalling >RhoA Kinase (Rock) and p90 Ribosomal S6 Kinase (p90Rsk) phosphorylation of the sodium hydrogen exchanger (NHE1) is required for lysophosphatidic acid-induced transport, cytoskeletal organization and migration
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RhoA Kinase (Rock) and p90 Ribosomal S6 Kinase (p90Rsk) phosphorylation of the sodium hydrogen exchanger (NHE1) is required for lysophosphatidic acid-induced transport, cytoskeletal organization and migration

机译：RhoA激酶（Rock）和p90核糖体S6激酶（p90Rsk）的磷酸氢钠（NHE1）磷酸化是溶血磷脂酸诱导的转运，细胞骨架组织和迁移所必需的

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摘要

The sodium hydrogen exchanger isoform one (NHE1) plays a critical role coordinating asymmetric events at the leading edge of migrating cells and is regulated by a number of phosphorylation events influencing both the ion transport and cytoskeletal anchoring required for directed migration. Lysophosphatidic add (LPA) activation of RhoA kinase (Rock) and the Ras-ERK growth factor pathway induces cytoskeletal reorganization, activates NHE1 and induces an increase in cell motility. We report that both Rock land II stoichiometrically phosphorylate NHE1 at threonine 653 in vitro using mass spectrometry and reconstituted kinase assays. In fibroblasts expressing NHE1 alanine mutants for either Rock (T653A) or ribosomal S6 kinase (Rsk; S703A) we show that each site is partially responsible for the LPA-induced increase in transport activity while NHE1 phosphorylation by either Rock or Rsk at their respective site is sufficient for LPA stimulated stress fiber formation and migration. Furthermore, mutation of either T653 or S703 leads to a higher basal pH level and a significantly higher proliferation rate. Our results identify the direct phosphorylation of NHE1 by Rock and suggest that both RhoA and Ras pathways mediate NHE1-dependent ion transport and migration in fibroblasts. (C) 2015 Elsevier Inc. All rights reserved.
机译：氢氢交换异构体一（NHE1）在协调迁移细胞前沿的不对称事件中起着关键作用，并受许多影响定向迁移所需的离子迁移和细胞骨架锚定的磷酸化事件的调节。 RhoA激酶（Rock）和Ras-ERK生长因子途径的溶血磷脂添加（LPA）激活诱导细胞骨架重组，激活NHE1并诱导细胞运动性增加。我们报告说，这两个Rock land II在质谱法和重组激酶测定中均在体外在苏氨酸653上化学计量地磷酸化了NHE1。在表达针对Rock（T653A）或核糖体S6激酶（Rsk; S703A）的NHE1丙氨酸突变体的成纤维细胞中，我们显示，每个位点都部分负责LPA诱导的转运活性增加，而Rock或Rsk在其各自位点使NHE1磷酸化LPA刺激的应力纤维形成和迁移就足够了。此外，T653或S703的突变会导致更高的基础pH值和明显更高的增殖率。我们的研究结果确定了Rock对NHE1的直接磷酸化作用，并表明RhoA和Ras途径均可介导NHE1依赖性离子在成纤维细胞中的迁移和迁移。 （C）2015 Elsevier Inc.保留所有权利。

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