FMNL2 and-3 regulate Golgi architecture and anterograde transport downstream of Cdc42
FMNL2 and-3 regulate Golgi architecture and anterograde transport downstream of Cdc42
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DOI：
10.1038/s41598-017-09952-1
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发表时间：
2017-08-29
影响因子：
4.6
通讯作者：
Rottner, Klemens
中科院分区：
文献类型：
--
作者：
Kage, Frieda;Steffen, Anika;Rottner, Klemens
rho家族小GTPase Cdc42定位于质膜和高尔基复体，除了突出和迁移外，还参与囊泡运输、内胞和胞外分泌以及细胞极性的建立和/或维持。formin家族成员FMNL2和-3是在迁移和侵袭过程中调节细胞边缘突起的肌动蛋白组装因子。在这里，我们报道这些形成蛋白在高尔基体中积累和发挥作用。与板足相反，高尔基体靶向这些蛋白需要它们的n端肉豆肉酰化和与Cdc42的相互作用。此外，FMNL2或-3的高尔基关联诱导了高尔基体周围可检测到生殖器素的肌动蛋白网络。重要的是，通过RNAi或CRISPR/ cas9介导的基因缺失对FMNL2/3形成蛋白的功能干扰不可避免地在不同细胞系中诱导高尔基体断裂。此外，缺乏这些蛋白质会导致核内体增大，以及成熟缺陷和/或分选为晚期核内体和溶酶体。FMNL2/3的耗损也影响VSV-G从高尔基体向质膜的顺行运输，与Cdc42（最近被发现通过货物分拣和载体形成来调节通过高尔基体的顺行运输）一致。因此，我们的数据将FMNL2/3形成蛋白与Cdc42在高尔基体顺行运输中的肌动蛋白组装依赖功能联系起来。
The Rho-family small GTPase Cdc42 localizes at plasma membrane and Golgi complex and aside from protrusion and migration operates in vesicle trafficking, endo- and exocytosis as well as establishment and/or maintenance of cell polarity. The formin family members FMNL2 and -3 are actin assembly factors established to regulate cell edge protrusion during migration and invasion. Here we report these formins to additionally accumulate and function at the Golgi apparatus. As opposed to lamellipodia, Golgi targeting of these proteins required both their N-terminal myristoylation and the interaction with Cdc42. Moreover, Golgi association of FMNL2 or -3 induced a phalloidin-detectable actin meshwork around the Golgi. Importantly, functional interference with FMNL2/3 formins by RNAi or CRISPR/Cas9-mediated gene deletion invariably induced Golgi fragmentation in different cell lines. Furthermore, absence of these proteins led to enlargement of endosomes as well as defective maturation and/or sorting into late endosomes and lysosomes. In line with Cdc42 - recently established to regulate anterograde transport through the Golgi by cargo sorting and carrier formation - FMNL2/3 depletion also affected anterograde trafficking of VSV-G from the Golgi to the plasma membrane. Our data thus link FMNL2/3 formins to actin assembly-dependent functions of Cdc42 in anterograde transport through the Golgi apparatus.