R-Ras regulates migration through an interaction with filamin A in melanoma cells.
R-Ras regulates migration through an interaction with filamin A in melanoma cells.
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DOI：
10.1371/journal.pone.0011269
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发表时间：
2010-06-23
期刊：
影响因子：
3.7
通讯作者：
Matter ML
中科院分区：
文献类型：
--
作者：
Gawecka JE;Griffiths GS;Ek-Rylander B;Ramos JW;Matter ML
肿瘤微环境中细胞粘附和迁移的变化是转移开始和进展的关键。R-Ras是调节细胞在细胞外基质上粘附和迁移的几种小GTP酶之一，但其机制尚未完全阐明。使用酵母双杂交方法，我们试图确定新的R-Ras结合蛋白，可能介导其对整合素的影响。我们确定细丝蛋白A（FLNa）作为候选相互作用蛋白。FLNa是一种肌动蛋白结合支架蛋白，也与整联蛋白β1、β2和β7尾结合，并与包括细胞迁移在内的多种细胞过程相关。事实上，M2黑色素瘤细胞需要FLNa用于运动。我们进一步表明，R-Ras和FLNa相互作用的免疫共沉淀和下拉测定。FLNa重复序列3（FLNaΔ3）的缺失消除了这种相互作用。在M2黑色素瘤细胞中，活性R-Ras与FLNa共定位，但不与缺乏重复序列3的FLNa共定位。因此，活化的R-Ras结合FLNa的重复序列3。这种相互作用的功能结果是，活性R-Ras和FLNa协调增加细胞迁移。相反，R-Ras和FLNaΔ3的共表达对迁移的影响显著降低。虽然有整合素活化和纤连蛋白基质组装的增强，细胞粘附没有改变。最后，内源性R-Ras的siRNA敲低损害FLNa依赖性纤连蛋白基质组装。这些数据支持一种模型，其中R-Ras与FLNa功能性相关，从而调节整合素依赖性迁移。因此，在黑色素瘤细胞中，R-Ras和FLNa可以通过增强细胞迁移来协同促进转移。
Changes in cell adhesion and migration in the tumor microenvironment are key in the initiation and progression of metastasis. R-Ras is one of several small GTPases that regulate cell adhesion and migration on the extracellular matrix, however the mechanism has not been completely elucidated. Using a yeast two-hybrid approach we sought to identify novel R-Ras binding proteins that might mediate its effects on integrins. We identified Filamin A (FLNa) as a candidate interacting protein. FLNa is an actin-binding scaffold protein that also binds to integrin β1, β2 and β7 tails and is associated with diverse cell processes including cell migration. Indeed, M2 melanoma cells require FLNa for motility. We further show that R-Ras and FLNa interact in co-immunoprecipitations and pull-down assays. Deletion of FLNa repeat 3 (FLNaΔ3) abrogated this interaction. In M2 melanoma cells active R-Ras co-localized with FLNa but did not co-localize with FLNa lacking repeat 3. Thus, activated R-Ras binds repeat 3 of FLNa. The functional consequence of this interaction was that active R-Ras and FLNa coordinately increased cell migration. In contrast, co-expression of R-Ras and FLNaΔ3 had a significantly reduced effect on migration. While there was enhancement of integrin activation and fibronectin matrix assembly, cell adhesion was not altered. Finally, siRNA knockdown of endogenous R-Ras impaired FLNa-dependent fibronectin matrix assembly. These data support a model in which R-Ras functionally associates with FLNa and thereby regulates integrin-dependent migration. Thus in melanoma cells R-Ras and FLNa may cooperatively promote metastasis by enhancing cell migration.
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DOI：
10.1083/jcb.145.5.1077
发表时间：
1999-05-31
期刊：
The Journal of cell biology
影响因子：
--
作者：
Keely PJ;Rusyn EV;Cox AD;Parise LV
通讯作者：
Parise LV
影响因子：
4.8
作者：
Green, J. Angelo;Berrier, Allison L.;Yamada, Kenneth M.
通讯作者：
Yamada, Kenneth M.
影响因子：
3.7
作者：
Baldassarre M;Razinia Z;Burande CF;Lamsoul I;Lutz PG;Calderwood DA
通讯作者：
Calderwood DA
影响因子：
8
作者：
Oertli, B;Han, J;Hughes, PE
通讯作者：
Hughes, PE
影响因子：
3.5
作者：
Cukier, I. Howard;Li, Yun;Lee, Jonathan M.
通讯作者：
Lee, Jonathan M.