Extraction of active RhoGTPases by RhoGDI regulates spatiotemporal patterning of RhoGTPases
Extraction of active RhoGTPases by RhoGDI regulates spatiotemporal patterning of RhoGTPases
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DOI：
10.7554/elife.50471
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发表时间：
2019-10-24
期刊：
影响因子：
7.7
通讯作者：
Bement, William M.
中科院分区：
文献类型：
--
作者：
Golding, Adriana E.;Visco, Ilaria;Bement, William M.
RhoGTP酶的特征在于作为膜相关的分子开关，其在活性的GTP结合状态和非活性的GDP结合状态之间循环。然而，90-95%的RhoGTP酶通过RhoGDI维持在可溶形式，其通常被视为失活RhoGTP酶的被动穿梭。我们目前对RhoGTPases的理解：RhoGDI动力学受到两个实验挑战的限制：体内RhoGTPases的直接可视化和体外循环的重建。我们开发了直接成像脊椎动物RhoGTPases在体内或脂质双层在体外的方法。使用这些方法，我们鉴定了与膜相关的活性和非活性RhoGTPases的库，发现RhoGDI可以提取非活性和活性RhoGTPases，并且发现活性RhoGTPases的提取有助于它们在细胞伤口周围的空间调节。这些结果表明，RhoGDI通过从质膜上去除活性RhoGTPases直接有助于RhoGTPases的时空模式。
The RhoGTPases are characterized as membrane-associated molecular switches that cycle between active, GTP-bound and inactive, GDP-bound states. However, 90-95% of RhoGTPases are maintained in a soluble form by RhoGDI, which is generally viewed as a passive shuttle for inactive RhoGTPases. Our current understanding of RhoGTPase:RhoGDI dynamics has been limited by two experimental challenges: direct visualization of the RhoGTPases in vivo and reconstitution of the cycle in vitro. We developed methods to directly image vertebrate RhoGTPases in vivo or on lipid bilayers in vitro. Using these methods, we identified pools of active and inactive RhoGTPase associated with the membrane, found that RhoGDI can extract both inactive and active RhoGTPases, and found that extraction of active RhoGTPase contributes to their spatial regulation around cell wounds. These results indicate that RhoGDI directly contributes to the spatiotemporal patterning of RhoGTPases by removing active RhoGTPases from the plasma membrane.