TY - JOUR TI - Extraction of active RhoGTPases by RhoGDI regulates spatiotemporal patterning of RhoGTPases AU - Golding, Adriana E AU - Visco, Ilaria AU - Bieling, Peter AU - Bement, William M A2 - Chernoff, Jonathan A2 - Akhmanova, Anna A2 - Chernoff, Jonathan VL - 8 PY - 2019 DA - 2019/10/24 SP - e50471 C1 - eLife 2019;8:e50471 DO - 10.7554/eLife.50471 UR - https://doi.org/10.7554/eLife.50471 AB - 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. KW - RhoGTPase KW - RhoGDI KW - Cdc42 KW - Rho JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -