MARCKS-related protein regulates cytoskeletal organization at cell-cell and cell-substrate contacts in epithelial cells
MARCKS-related protein regulates cytoskeletal organization at cell-cell and cell-substrate contacts in epithelial cells
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DOI：
10.1242/jcs.210237
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发表时间：
2018-02-01
影响因子：
4
通讯作者：
Anderson, James M.
中科院分区：
文献类型：
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作者：
Van Itallie, Christina M.;Tietgens, Amber Jean;Anderson, James M.
用干扰素-γ和TNF-α（IFN/TNF）处理上皮细胞导致细胞旁通透性增加。为了鉴定介导屏障破坏的相关蛋白质，我们对occludin进行了邻近依赖性生物素化（BioID），发现MARCKS相关蛋白（MRP;也称为MARCKSL 1）的标记在IFN/TNF给药后增加了20倍。GFP-MRP集中在细胞侧膜，其过表达增强了紧密连接屏障对细胞因子的生理反应。然而，MRP的缺失并没有消除细胞因子的反应，这表明MRP是不需要在闭塞素依赖性IFN/TNF的反应。相反，我们的研究结果揭示了MRP在上皮细胞中控制多种肌动蛋白结构的关键作用，可能是通过调节整合素信号传导。MRP敲除（KO）细胞中粘着斑组织和基底肌动蛋白应力纤维的变化让人想起FAK-KO细胞中观察到的变化。此外，我们发现MRP-KO细胞中细胞-细胞相互作用的改变与连接张力增加相关，表明MRP可能在局灶性粘附-粘附连接串扰中发挥作用。总之，我们的结果是一致的MRP在上皮细胞中的细胞接触的细胞骨架组织的关键作用。
Treatment of epithelial cells with interferon-gamma and TNF-alpha (IFN/TNF) results in increased paracellular permeability. To identify relevant proteins mediating barrier disruption, we performed proximity-dependent biotinylation (BioID) of occludin and found that tagging of MARCKS-related protein (MRP; also known as MARCKSL1) increased similar to 20-fold following IFN/TNF administration. GFP-MRP was focused at the lateral cell membrane and its overexpression potentiated the physiological response of the tight junction barrier to cytokines. However, deletion of MRP did not abrogate the cytokine responses, suggesting that MRP is not required in the occludin-dependent IFN/TNF response. Instead, our results reveal a key role for MRP in epithelial cells in control of multiple actin-based structures, likely by regulation of integrin signaling. Changes in focal adhesion organization and basal actin stress fibers in MRP-knockout (KO) cells were reminiscent of those seen in FAK-KO cells. In addition, we found alterations in cell-cell interactions in MRP-KO cells associated with increased junctional tension, suggesting that MRP may play a role in focal adhesion-adherens junction cross talk. Together, our results are consistent with a key role for MRP in cytoskeletal organization of cell contacts in epithelial cells.