- Title
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RAC1-Dependent ORAI1 Translocation to the Leading Edge Supports Lamellipodia Formation and Directional Persistence
- Authors
- Lopez-Guerrero, A.M., Espinosa-Bermejo, N., Sanchez-Lopez, I., Macartney, T., Pascual-Caro, C., Orantos-Aguilera, Y., Rodriguez-Ruiz, L., Perez-Oliva, A.B., Mulero, V., Pozo-Guisado, E., Martin-Romero, F.J.
- Source
- Full text @ Sci. Rep.
Genetic ablation of ORAI1 reduced motility, directness, lamellipodia formation, and invasion of U2OS cells. |
EGF potentiated ORAI1 binding to CTTN, CYFIP1, and ARP2/3. |
Activation of RAC1 triggered the translocation of ORAI1 to CTTN-containing cell periphery. |
RAC1 regulated the dynamics of peripheral ORAI1-CTTN and ruffling. U2OS cells stably expressing Flag-RAC1 (wild-type), Flag-RAC1G12V or Flag-RAC1T17N, were transfected for the transient expression of ORAI1-GFP and mCherry-CTTN. Fluorescence was monitored in live cells in Leibovitz’s L-15 medium supplemented with 10% FBS. The ROI indicated by the arrow was selected to assess the variation of fluorescence intensity (right panels). Emission of fluorescence of ORAI1-GFP (green line) and mCherry-CTTN (red line) was recorded every 3 sec for 4 min. Full time-lapse sequences are shown as Supplementary Movies |
RAC1T17N blocked the transport of ORAI1 to the cell surface. |
RAC1 co-precipitated with ORAI1 upon stimulation with EGF. |
Inhibition of RAC1 impaired cell motility and reduced co-precipitation of ORAI1 with markers of the leading edge. |
Inhibition of RAC1 reduced surface ORAI1 and SOCE. |