Cell migration is critical for several physiological and pathophysiological processes. It depends on the coordinated action of kinases, phosphatases, Rho-GTPases proteins, and Ca²⁺ signaling. Interestingly, ubiquitination events have emerged as regulatory elements of migration. Thus, the role of proteins involved in ubiquitination processes could be relevant to a complete understanding of pro-migratory mechanisms. KCTD5 is a member of Potassium Channel Tetramerization Domain (KCTD) proteins that have been proposed as a putative adaptor for Cullin3-E3 ubiquitin ligase and a novel regulatory protein of TRPM4 channels. Here, we study whether KCTD5 participates in cell migration-associated mechanisms, such as focal adhesion dynamics and cellular spreading. Our results show that KCTD5 CRISPR/Cas9- and shRNA-based depletion in B16-F10 cells promoted an increase in cell migration and cell spreading, and a decrease in the focal adhesion area, consistent with an increased focal adhesion disassembly rate. The expression of a dominant-negative mutant of Rho-GTPases Rac1 precluded the KCTD5 depletion-induced increase in cell spreading. Additionally, KCTD5 silencing decreased the serum-induced Ca²⁺ response, and the reversion of this with ionomycin abolished the KCTD5 knockdown-induced decrease in focal adhesion size. Together, these data suggest that KCTD5 acts as a regulator of cell migration by modulating cell spreading and focal adhesion dynamics through Rac1 activity and Ca²⁺ signaling, respectively.

中文翻译：

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K +通道四聚化域5（KCTD5）蛋白通过调节Ca 2+信号传导和Rac1活性来调节细胞迁移，局部黏附动力学和扩散。

细胞迁移对于几个生理和病理生理过程至关重要。它取决于激酶，磷酸酶，Rho-GTPases蛋白和Ca ²⁺的协同作用。信号。有趣的是，泛素化事件已成为移民的监管要素。因此，参与泛素化过程的蛋白质的作用可能与对促迁移机制的完整理解有关。KCTD5是钾通道四聚化域（KCTD）蛋白的成员，该蛋白已被提议作为Cullin3-E3泛素连接酶和TRPM4通道的新型调节蛋白的假定衔接子。在这里，我们研究KCTD5是否参与细胞迁移相关的机制，如粘着斑动态和细胞扩散。我们的结果表明，B16-F10细胞中基于KCTD5 CRISPR / Cas9-和shRNA的耗竭促进细胞迁移和细胞扩散的增加​​，以及粘着斑面积的减少，这与粘着斑的拆卸率增加相一致。Rho-GTPases Rac1的显性负突变体的表达排除了KCTD5耗竭诱导的细胞扩散增加。此外，KCTD5沉默降低了血清诱导的钙²⁺反应，并用离子霉素逆转，消除了KCTD5敲低引起的粘着斑大小的减少。总之，这些数据表明，KCTD5通过分别通过Rac1活性和Ca ²⁺信号传导调节细胞扩散和粘着斑动态来充当细胞迁移的调节剂。