Loss of Ena/VASP interferes with lamellipodium architecture, motility and integrin-dependent adhesion
Loss of Ena/VASP interferes with lamellipodium architecture, motility and integrin-dependent adhesion
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DOI：
10.7554/elife.55351
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发表时间：
2020-05-11
期刊：
影响因子：
7.7
通讯作者：
Faix, Jan
中科院分区：
文献类型：
--
作者：
Damiano-Guercio, Julia;Kurzawa, Laetitia;Faix, Jan
细胞迁移需要肌动蛋白丝的网络和束，分别称为板状伪足和微刺或丝状伪足，以及局灶性粘附，所有这些都招募Ena/VASP家族成员迄今被认为拮抗有效的细胞运动。然而，我们发现这些蛋白质在不同的鼠细胞系中作为迁移的正调节因子。CRISPR/Cas9介导的Ena/VASP蛋白的损失减少了片状伪足肌动蛋白组装并扰乱了片状伪足结构，如通过改变的网络几何形状以及伴随异常Arp 2/3复合物和异二聚体加帽蛋白积累的细丝长度和数量的减少所证明的。Ena/VASP功能的丧失也废除了通常嵌入板状伪足中的微刺的形成，但不包括能够在没有板状伪足的情况下发出的丝状伪足。Ena/VASP缺陷也损害了整合素介导的粘附，伴随着通过这些结构施加的牵引力减少。因此，我们的数据揭示了这些肌动蛋白聚合酶的新Ena/VASP功能，这与它们促进细胞迁移完全一致。
Cell migration entails networks and bundles of actin filaments termed lamellipodia and microspikes or filopodia, respectively, as well as focal adhesions, all of which recruit Ena/VASP family members hitherto thought to antagonize efficient cell motility. However, we find these proteins to act as positive regulators of migration in different murine cell lines. CRISPR/Cas9-mediated loss of Ena/VASP proteins reduced lamellipodial actin assembly and perturbed lamellipodial architecture, as evidenced by changed network geometry as well as reduction of filament length and number that was accompanied by abnormal Arp2/3 complex and heterodimeric capping protein accumulation. Loss of Ena/VASP function also abolished the formation of microspikes normally embedded in lamellipodia, but not of filopodia capable of emanating without lamellipodia. Ena/VASP-deficiency also impaired integrin-mediated adhesion accompanied by reduced traction forces exerted through these structures. Our data thus uncover novel Ena/VASP functions of these actin polymerases that are fully consistent with their promotion of cell migration.