Paxillin and Hic-5 interaction with vinculin is differentially regulated by Rac1 and RhoA.
Paxillin and Hic-5 interaction with vinculin is differentially regulated by Rac1 and RhoA.
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DOI：
10.1371/journal.pone.0037990
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Turner CE
中科院分区：
文献类型：
--
作者：
Deakin NO;Ballestrem C;Turner CE
细胞迁移对于生物体的发育和维持以及包括癌症转移在内的多种病理过程都是至关重要的。RhoGTP酶rac1和RhoA调节细胞突起、细胞-细胞外基质(ECM)相互作用和力传导，对细胞迁移是不可或缺的。然而，它们在细胞内分子水平上的活动的后果仍然不确定。利用FRET、FRAP和生化分析相结合的方法，我们发现粘着斑蛋白vinculin和paxlin以及密切相关的家族成员Hic-5之间的相互作用在空间上和相互作用中受到rac1和RhoA活性的调节。当rac1和RhoA被激活时，其活性构象中的vinculin分别与粘连中的paxlin或Hic-5相互作用，而在rac1被抑制后，非活性vinculin与膜中的paxlin相互作用。此外，Rac1还特异性地调节粘连中的巴西林及其结合伙伴和F-肌动蛋白相互作用蛋白Actopaxin(α-Parvin)的动态。此外，对蛋白质的FRET分析：3D基质中形成的细胞粘连中的蛋白质相互作用表明，与2D系统相比，vinculin优先与Hic-5相互作用。这项研究通过首次描述RhoGTPase协调蛋白：细胞微环境中的蛋白质相互作用，为2D和3D基质中细胞-ECM黏附的复杂性提供了新的见解。这些数据确定了paxlin和Hic-5在rac1和RhoA依赖的细胞黏附形成和成熟过程中的不同角色；这些过程对于高效的细胞迁移至关重要。
Cell migration is of paramount importance to organism development and maintenance as well as multiple pathological processes, including cancer metastasis. The RhoGTPases Rac1 and RhoA are indispensable for cell migration as they regulate cell protrusion, cell-extracellular matrix (ECM) interactions and force transduction. However, the consequences of their activity at a molecular level within the cell remain undetermined. Using a combination of FRET, FRAP and biochemical analyses we show that the interactions between the focal adhesion proteins vinculin and paxillin, as well as the closely related family member Hic-5 are spatially and reciprocally regulated by the activity of Rac1 and RhoA. Vinculin in its active conformation interacts with either paxillin or Hic-5 in adhesions in response to Rac1 and RhoA activation respectively, while inactive vinculin interacts with paxillin in the membrane following Rac1 inhibition. Additionally, Rac1 specifically regulates the dynamics of paxillin as well as its binding partner and F-actin interacting protein actopaxin (α-parvin) in adhesions. Furthermore, FRET analysis of protein:protein interactions within cell adhesions formed in 3D matrices revealed that, in contrast to 2D systems vinculin interacts preferentially with Hic-5. This study provides new insight into the complexity of cell-ECM adhesions in both 2D and 3D matrices by providing the first description of RhoGTPase-coordinated protein:protein interactions in a cellular microenvironment. These data identify discrete roles for paxillin and Hic-5 in Rac1 and RhoA-dependent cell adhesion formation and maturation; processes essential for productive cell migration.
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影响因子：
3.3
作者：
Deakin NO;Turner CE
通讯作者：
Turner CE
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作者：
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56.9
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