Ezrin/Radixin/Moesin proteins and flotillins cooperate to promote uropod formation in T cells.
Ezrin/Radixin/Moesin proteins and flotillins cooperate to promote uropod formation in T cells.
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DOI：
10.3389/fimmu.2013.00084
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发表时间：
2013
影响因子：
7.3
通讯作者：
Niggli V
中科院分区：
文献类型：
--
作者：
Martinelli S;Chen EJ;Clarke F;Lyck R;Affentranger S;Burkhardt JK;Niggli V
T细胞尾足富含特异性蛋白质，包括粘附受体如P-选择素糖蛋白配体-1（PSGL-1）、脂筏相关蛋白如漂浮蛋白和与富含胆固醇的筏结构域相关的埃兹蛋白/根蛋白/膜突蛋白（ERM）蛋白，并将粘附受体锚在肌动蛋白细胞骨架上。使用显性突变体和siRNA技术，我们已经测试了这些蛋白质之间的相互作用及其在塑造T细胞尾足中的作用。野生型（WT）ezrin-EGFP的表达未能影响人T细胞的形态或趋化因子诱导的PSGL-1和flotillin-1和-2的尾足募集。相反，组成型活性T567 D ezrin-EGFP的表达诱导了一些转染的T细胞中的能动的极化表型，即使在没有趋化因子的情况下。这些细胞的特点是F-肌动蛋白丰富的褶皱在前面和尾足富集PSGL-1和flotillins。T567 D ezrin-EGFP本身在极化T细胞的后部强烈富集。T567 D ezrin-EGFP诱导的尾足形成是肌动蛋白依赖性的，因为它通过抑制Rho激酶或肌球蛋白II而减弱，并且通过破坏肌动蛋白丝而消除。虽然组成型活性ezrin的表达增强细胞极性，但ezrin的显性负性缺失突变体1-310 ezrin-EGFP的表达显著降低了趋化因子SDF-1诱导的尾足形成、T细胞的前-尾极性以及PSGL-1和flotillin的加帽。用WT或T567 D ezrin转染T细胞不影响趋化因子介导的趋化性，而1-310 ezrin显著损害自发的2D迁移和趋化性。siRNA介导的小鼠T细胞中flotillin的下调减弱了膜突蛋白加帽和尾足形成，表明ERM蛋白和flotillin在尾足形成中合作。总之，我们的研究结果表明，激活的ERM蛋白的功能与flotillins，以促进有效的趋化性T细胞的结构uropod迁移T细胞。
T cell uropods are enriched in specific proteins including adhesion receptors such as P-selectin glycoprotein ligand-1 (PSGL-1), lipid raft-associated proteins such as flotillins and ezrin/radixin/moesin (ERM) proteins which associate with cholesterol-rich raft domains and anchor adhesion receptors to the actin cytoskeleton. Using dominant mutants and siRNA technology we have tested the interactions among these proteins and their role in shaping the T cell uropod. Expression of wild type (WT) ezrin-EGFP failed to affect the morphology of human T cells or chemokine-induced uropod recruitment of PSGL-1 and flotillin-1 and -2. In contrast, expression of constitutively active T567D ezrin-EGFP induced a motile, polarized phenotype in some of the transfected T cells, even in the absence of chemokine. These cells featured F-actin-rich ruffles in the front and uropod enrichment of PSGL-1 and flotillins. T567D ezrin-EGFP was itself strongly enriched in the rear of the polarized T cells. Uropod formation induced by T567D ezrin-EGFP was actin-dependent as it was attenuated by inhibition of Rho-kinase or myosin II, and abolished by disruption of actin filaments. While expression of constitutively active ezrin enhanced cell polarity, expression of a dominant-negative deletion mutant of ezrin, 1–310 ezrin-EGFP, markedly reduced uropod formation induced by the chemokine SDF-1, T cell front-tail polarity, and capping of PSGL-1 and flotillins. Transfection of T cells with WT or T567D ezrin did not affect chemokine-mediated chemotaxis whereas 1–310 ezrin significantly impaired spontaneous 2D migration and chemotaxis. siRNA-mediated downregulation of flotillins in murine T cells attenuated moesin capping and uropod formation, indicating that ERM proteins and flotillins cooperate in uropod formation. In summary, our results indicate that activated ERM proteins function together with flotillins to promote efficient chemotaxis of T cells by structuring the uropod of migrating T cells.
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DOI：
10.1083/jcb.201005140
发表时间：
2010-11-15
期刊：
The Journal of cell biology
影响因子：
--
作者：
Ludwig A;Otto GP;Riento K;Hams E;Fallon PG;Nichols BJ
通讯作者：
Nichols BJ
影响因子：
3.7
作者：
Chen EJ;Shaffer MH;Williamson EK;Huang Y;Burkhardt JK
通讯作者：
Burkhardt JK
磷酸肌醇的结合和磷酸化在ezrin的激活机理中依次起作用。
影响因子：
7.8
作者：
Fievet, BT;Gautreau, A;Roy, C;Del Maestro, L;Mangeat, P;Louvard, D;Arpin, M
通讯作者：
Arpin, M
DOI：
10.1083/jcb.201201018
发表时间：
2012-06-11
期刊：
The Journal of cell biology
影响因子：
--
作者：
Babich A;Li S;O'Connor RS;Milone MC;Freedman BD;Burkhardt JK
通讯作者：
Burkhardt JK
影响因子：
20.3
作者：
Millán, J;Montoya, MC;Alonso, MA
通讯作者：
Alonso, MA