The formin-homology-domain-containing protein FHOD1 enhances cell migration
The formin-homology-domain-containing protein FHOD1 enhances cell migration
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DOI：
10.1242/jcs.00386
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发表时间：
2003-05-01
影响因子：
4
通讯作者：
Westendorf, JJ
中科院分区：
文献类型：
--
作者：
Koka, S;Neudauer, CL;Westendorf, JJ
含有Forin同源结构域的蛋白与Rho家族GTP酶相互作用，调节肌动蛋白的细胞骨架组织和基因转录。FHOD1是这个家族的成员之一，与rac1相互作用，并从血清反应元件诱导转录。在这项研究中，我们研究了FHOD1表达对哺乳动物细胞中细胞骨架组织和功能的影响。FHOD1蛋白在WM35黑色素瘤细胞和NIH-3T3成纤维细胞中稳定表达。表达全长FHOD1的细胞与载体转染的细胞和表达截短的FHOD1(1-421)的细胞相比，表现出拉长的表型，这些细胞缺乏保守的FH1和FH2结构域。全长FHOD1与丝状肌动蛋白共同定位于细胞外围。瞬时表达C端FHOD1截断突变体(DeltaC，残基1-1010)的细胞显示出显著的应激纤维，该突变体缺乏自抑制蛋白质-蛋白质相互作用结构域。FHOD1(1-421)不诱导应激纤维，但以类似于全长蛋白的方式定位于膜褶皱，表明FH1和FH2结构域是应激纤维出现所必需的。依赖FHOD1DeltaC(1-1010)的应力纤维对显性负值的racN17、RhoA和ROCK抑制剂、C3转移酶和Y-27632敏感。稳定过表达全长FHOD1可促进WM35和NIH-3T3细胞分别向I型胶原和纤维连接蛋白迁移。表达FHOD1(1-421)的细胞迁移与对照细胞相似。整合素的表达和活化不受FHOD1表达的影响。此外，FHOD1的过表达不会改变黏附或迁移过程中整合素的使用。这些数据表明，FHOD1以不依赖整合素的方式与细胞骨架相互作用并调节细胞骨架的结构，并刺激细胞迁移。
Formin-homology-domain-containing proteins interact with Rho-family GTPases and regulate actin cytoskeleton organization and gene transcription. FHOD1 is a member of this family, interacts with Rac1 and induces transcription from the serum response element. In this study, we examined the effects of FHOD1 expression on cytoskeletal organization and function in mammalian cells. FHOD1 proteins were stably expressed in WM35 melanoma cells and NIH-3T3 fibroblasts. Cells expressing full-length FHOD1 demonstrated an elongated phenotype compared with vector-transfected cells and cells expressing a truncated FHOD1 (1-421) that lacks the conserved FH1 and FH2 domains. Full-length FHOD1 co-localized with filamentous actin at cell peripheries. Cells transiently expressing a C-terminal FHOD1 truncation mutant (DeltaC, residues 1-1010), which lacks an autoinhibitory protein-protein interaction domain, displayed prominent stress fibers. FHOD1 (1-421) did not induce stress fibers but localized to membrane ruffles in a manner similar to the full-length protein, indicating that the FH1 and FH2 domains are required for stress fiber appearance. FHOD1 DeltaC (1-1010)-dependent stress fibers were sensitive to dominant-negative RacN17 and the RhoA and ROCK inhibitors, C3 transferase and Y-27632. Stable overexpression of full-length FHOD1 enhanced the migration of WM35 and NIH-3T3 cells to type-I collagen and fibronectin, respectively. Cells expressing FHOD1 (1-421) migrated similar to control cells. Integrin expression and activation were not affected by FHOD1 expression. Moreover, FHOD1 overexpression did not alter integrin usage during adhesion or migration. These data demonstrate that FHOD1 interacts with and regulates the structure of the cytoskeleton and stimulates cell migration in an integrin-independent manner.