The microtubule-associated protein EB1 maintains cell polarity through activation of protein kinase C.
The microtubule-associated protein EB1 maintains cell polarity through activation of protein kinase C.
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DOI：
10.1016/j.bbrc.2011.11.056
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发表时间：
2012-01-06
影响因子：
3.1
通讯作者：
Cain JM
中科院分区：
文献类型：
--
作者：
Schober JM;Kwon G;Jayne D;Cain JM
微管的正端靶向细胞皮质以调节肌动蛋白突起动力学和极性，但很少有人知道耦合相互作用的分子机制。EB 1蛋白与微管的正端结合，将EB 1置于理想的空间位置以介导微管-肌动蛋白串扰。本研究的目的是进一步了解EB 1依赖的细胞极性和运动性的细胞内信号转导。使用短发针RNA干扰消除B16 F10小鼠黑色素瘤细胞中的EB 1蛋白。进行相关活细胞免疫荧光显微镜检查以确定WAVE 2和IQGAP 1在突出边缘与缩回边缘的定位。EB 1敲低导致WAVE 2和IQGAP 1的亚细胞分离不良，并且总体定位降低。PKC的激活纠正了WAVE 2和IQGAP 1定位、细胞扩散和细胞形状的缺陷，达到对照细胞中观察到的水平，但没有纠正细胞迁移的缺陷。与这些发现一致，在EB 1敲低细胞中观察到PKC磷酸化降低。这些发现支持了一个模型，即EB 1蛋白通过涉及PKC的信号通路将微管与肌动蛋白突起和细胞极性联系起来。
The plus-ends of microtubules target the cell cortex to modulate actin protrusion dynamics and polarity, but little is known of the molecular mechanism that couples the interaction. EB1 protein associates with the plus-ends of microtubules, placing EB1 in an ideal spatial position to mediate microtubule-actin cross talk. The objective of the current study was to further understand intracellular signaling involved in EB1-dependent cell polarity and motility. B16F10 mouse melanoma cells were depleted of EB1 protein using short hair-pin RNA interference. Correlative live cell-immunofluorescence microscopy was performed to determine localization of WAVE2 and IQGAP1 to protruding versus retracting edges. EB1 knock down caused poor subcellular separation of WAVE2 and IQGAP1, and overall decreased localization. Activation of PKC corrected defects in WAVE2 and IQGAP1 localization, cell spreading and cell shape to levels observed in control cells, but did not correct defects in cell migration. Consistent with these findings, decreased PKC phosphorylation was observed in EB1 knock down cells. These findings support a model where EB1 protein links microtubules to actin protrusion and cell polarity through signaling pathways involving PKC.
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DOI：
10.1083/jcb.149.4.761
发表时间：
2000-05-15
期刊：
The Journal of cell biology
影响因子：
--
作者：
Tirnauer JS;Bierer BE
通讯作者：
Bierer BE
DOI：
10.1083/jcb.200603013
发表时间：
2006-09-11
期刊：
The Journal of cell biology
影响因子：
--
作者：
Vignjevic D;Kojima S;Aratyn Y;Danciu O;Svitkina T;Borisy GG
通讯作者：
Borisy GG
影响因子：
4
作者：
Schober, Joseph M.;Komarova, Yulia A.;Borisy, Gary G.
通讯作者：
Borisy, Gary G.
影响因子：
2.7
作者：
Kojima, SI;Vignjevic, D;Borisy, GG
通讯作者：
Borisy, GG
影响因子：
2.9
作者：
Slater, SJ;Seiz, JL;Stubbs, CD
通讯作者：
Stubbs, CD