The Carboxy-Terminal Tail of Connexin43 Gap Junction Protein Is Sufficient to Mediate Cytoskeleton Changes in Human Glioma Cells
The Carboxy-Terminal Tail of Connexin43 Gap Junction Protein Is Sufficient to Mediate Cytoskeleton Changes in Human Glioma Cells
复制标题
DOI：
10.1002/jcb.22554
复制
发表时间：
2010-06-01
影响因子：
4
通讯作者：
Sin, Wun-Chey
中科院分区：
文献类型：
--
作者：
Crespin, Sophie;Bechberger, John;Sin, Wun-Chey
缝隙连接蛋白43（Cx43）是差距连接蛋白家族中广泛表达的一个成员，它通过允许胞质物质的交换来介导缝隙连接细胞间通讯（GJIC）。先前的研究已经使用在胞质尾（C-尾）处截短的Cx43来证明C-尾对于调节细胞生长和运动是必不可少的。因此，我们的研究的目的是描绘各自的作用，截断的Cx43和C-尾介导的Cx43依赖的信号。将表达蛋白质的通道部分的截短的Cx43（TrCx 43，氨基酸1-242）和仅包含来自氨基酸243的C-尾的构建体（243 Cx43）转导到LN 18人神经胶质瘤细胞中。我们的研究结果表明，Cx43抑制生长的能力是独立的GJIC评估染料转移，但依赖于刚性细胞外基质的存在。我们进一步证明，单独的C-尾足以促进运动。令人惊讶的是，Cx43也能够在C尾不存在的情况下增加迁移，这表明Cx43利用至少两种不同的信号传导机制来影响运动性。最后，我们使用延时成像来检查迁移细胞的行为，并且很明显，C-尾与在模拟或表达TrCx 43的LN 18细胞中未观察到的基于板状伪足的迁移相关。我们的研究首次表明，一个自由的C-尾是足以诱导Cx43依赖的细胞形态学的变化，Cx43信号与肌动蛋白细胞骨架的调节。J.细胞。110：589-597，2010. (C)2010 Wiley-Liss，Inc.
Connexin43 (Cx43) is a ubiquitously expressed member of the gap junction protein family that mediates gap junction intercellular communication (GJIC) by allowing exchange of cytosolic materials. Previous studies have used Cx43 truncated at the cytoplasmic tail (C-tail) to demonstrate that the C-tail is essential to regulate cell growth and motility. Therefore, the aim of our study was to delineate the respective role of the truncated Cx43 and the C-tail in mediating Cx43-dependent signaling. A truncated Cx43 expressing the channel part of the protein (TrCx43, amino acid 1-242) and a construct encompassing only the C-tail from amino acid 243 (243Cx43) were transduced into LN18 human glioma cells. Our results showed that the ability of Cx43 to suppress growth was independent of GJIC as assessed by dye transfer, but was dependent on the presence of a rigid extracellular matrix. We further demonstrated that the C-tail alone is sufficient to promote motility. Surprisingly, Cx43 is also able to increase migration in the absence of the C-tail, suggesting the presence of at least two distinct signaling mechanisms utilized by Cx43 to affect motility. Finally, we used time-lapse imaging to examine the behavior of migrating cells and it was apparent that the C-tail was associated with a lamellipodia-based migration not observed in either mock or TrCx43 expressing LN18 cells. Our study shows for the first time that a free C-tail is sufficient to induce Cx43-dependent changes in cell morphology and that Cx43 signaling is linked to the regulation of the actin cytoskeleton. J. Cell. Biochem. 110: 589-597, 2010. (C) 2010 Wiley-Liss, Inc.