Il-Young Hwang, Ji Sung Kim, Kathleen A. Harrison, Chung Park, Chong Shan Shi, John H. Kehrl
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引用次数: 0
摘要
配体参与的趋化因子受体会触发异三聚体 Gαi 蛋白的核苷酸交换,从而刺激细胞骨架重组和细胞极性变化。为了更好地了解导致这些细胞变化的信号事件,我们重点研究了小鼠脾脏B细胞与趋化因子受体CXCR5接触后F-肌动蛋白动态的早期变化。在接触 CXCR5 配体 CXCL13 的 10 秒钟内,出现了三维片状伪足和富含 F-肌动蛋白的脊。瞬时 F-肌动蛋白的增加取决于 Gαi2/3 信号传导、PI3K/AKT 通路、ERK 激活、磷脂酶 C 活性以及由 Dock2(细胞分裂驱动因子 2)介导的 Rac1/2 激活。免疫印迹分析发现,激酶 WNK1(无赖氨酸激酶 1)是一种潜在的早期 AKT 效应因子。用特异性 WNK 抑制剂处理 B 细胞会破坏 F-肌动蛋白动力学,损害 B 细胞的极性、运动性和趋化性。通过对 Wnk1 进行 CRISPR-Cas9 基因编辑,在小鼠 B 细胞系中模拟了这些变化,这也表明 WNK1 对 B 细胞增殖有贡献。服用单剂量 WNK 抑制剂可短暂降低活体小鼠淋巴结中 B 细胞的运动性和极性。这些结果表明,WNK1 信号传导可维持 B 细胞对 CXCL13 的反应性,并表明药理抑制 WNK1 可能会影响体液免疫,而 WNK1 参与癌症进展和血压调节。
Chemokine-mediated F-actin dynamics, polarity, and migration in B lymphocytes depend on WNK1 signaling
Ligand-engaged chemokine receptors trigger nucleotide exchange in heterotrimeric Gαi proteins, which stimulates cytoskeletal reorganization and cell polarity changes. To better understand the signaling events responsible for these cellular changes, we focused on early changes in F-actin dynamics after engagement of the chemokine receptor CXCR5 in murine splenic B cells. Within 10 seconds of exposure to the CXCR5 ligand CXCL13, three-dimensional lamellar-like pseudopods and F-actin–rich ridges appeared. The transient F-actin increase depended on Gαi2/3 signaling, the PI3K/AKT pathway, ERK activation, phospholipase C activity, and Rac1/2 activation mediated by Dock2 (dedicator of cytokinesis 2). Immunoblot analyses identified the kinase WNK1 (with no lysine kinase 1) as a potential early AKT effector. Treating B cells with specific WNK inhibitors disrupted F-actin dynamics and impaired B cell polarity, motility, and chemotaxis. These changes were mimicked in a murine B cell line by CRISPR-Cas9 gene editing of Wnk1, which also suggested that WNK1 contributed to B cell proliferation. Administration of a single dose of a WNK inhibitor transiently reduced B cell motility and polarity in the lymph nodes of live mice. These results indicate that WNK1 signaling maintains B cell responsiveness to CXCL13 and suggest that pharmacological inhibition of WNK1, which is involved in cancer progression and blood pressure regulation, may affect humoral immunity.
期刊介绍:
"Science Signaling" is a reputable, peer-reviewed journal dedicated to the exploration of cell communication mechanisms, offering a comprehensive view of the intricate processes that govern cellular regulation. This journal, published weekly online by the American Association for the Advancement of Science (AAAS), is a go-to resource for the latest research in cell signaling and its various facets.
The journal's scope encompasses a broad range of topics, including the study of signaling networks, synthetic biology, systems biology, and the application of these findings in drug discovery. It also delves into the computational and modeling aspects of regulatory pathways, providing insights into how cells communicate and respond to their environment.
In addition to publishing full-length articles that report on groundbreaking research, "Science Signaling" also features reviews that synthesize current knowledge in the field, focus articles that highlight specific areas of interest, and editor-written highlights that draw attention to particularly significant studies. This mix of content ensures that the journal serves as a valuable resource for both researchers and professionals looking to stay abreast of the latest advancements in cell communication science.