TY - JOUR
T1 - A novel spatiotemporal RhoC activation pathway locally regulates cofilin activity at invadopodia
AU - Bravo-Cordero, Jose Javier
AU - Oser, Matthew
AU - Chen, Xiaoming
AU - Eddy, Robert
AU - Hodgson, Louis
AU - Condeelis, John
N1 - Funding Information:
We thank the Condeelis, Cox, and Segall laboratories for helpful discussions and the Analytical Imaging Facility (AIF) of the Gruss Lipper Biophotonics Center at Albert Einstein College of Medicine for technical help. We thank Geoffrey Perumal (AIF) for his technical help with transmission electron microscopy. We thank Vladislav Verkhusha (Albert Einstein College of Medicine) for providing pm-TagRFP-T-C1 and David D. Schlaepfer (University of California, San Diego) for providing p190RhoGEF-GFP. This work was funded by GM064346 (J.J.B.-C.), CA150344 (J.J.B.-C., M.O., R.E., J.C.), CA100324 (X.C.), GM093121 (J.J.B.-C., L.H.), and “Sinsheimer Foundation Young Investigator Award” (L.H.).
PY - 2011/4/26
Y1 - 2011/4/26
N2 - Background: RhoGTPases have been implicated in the regulation of cancer metastasis. Invasive carcinoma cells form invadopodia, F-actin-rich matrix-degrading protrusions that are thought to be important for tumor cell invasion and intravasation. Regulation of actin dynamics at invadopodial protrusions is crucial to drive invasion. This process requires the severing activity of cofilin to generate actin-free barbed ends. Previous work demonstrates that cofilin's severing activity is tightly regulated through multiple mechanisms, including regulation of cofilin serine phosphorylation by Rho GTPases. However, it is not known which Rho GTPase is involved in regulating cofilin's phosphorylation status at invadopodia. Results: We show here, for the first time, how RhoC activation is controlled at invadopodia and how this activation regulates cofilin phosphorylation to control cofilin's generation of actin-free barbed ends. Live-cell imaging of fluorescent RhoC biosensor reveals that RhoC activity is spatially confined to areas surrounding invadopodia. This spatiotemporal restriction of RhoC activity is controlled by "spatially distinct regulatory elements" that confine RhoC activation within this compartment. p190RhoGEF localizes around invadopodia to activate RhoC, whereas p190RhoGAP localizes inside invadopodia to deactivate the GTPase within the structure. RhoC activation enhances cofilin phosphorylation outside invadopodia. Conclusion: These results show how RhoC activity is spatially regulated at invadopodia by p190RhoGEF and p190RhoGAP. RhoC activation in areas surrounding invadopodia restricts cofilin activity to within the invadopodium core, resulting in a focused invadopodial protrusion. This mechanism likely enhances tumor cell invasion during metastasis.
AB - Background: RhoGTPases have been implicated in the regulation of cancer metastasis. Invasive carcinoma cells form invadopodia, F-actin-rich matrix-degrading protrusions that are thought to be important for tumor cell invasion and intravasation. Regulation of actin dynamics at invadopodial protrusions is crucial to drive invasion. This process requires the severing activity of cofilin to generate actin-free barbed ends. Previous work demonstrates that cofilin's severing activity is tightly regulated through multiple mechanisms, including regulation of cofilin serine phosphorylation by Rho GTPases. However, it is not known which Rho GTPase is involved in regulating cofilin's phosphorylation status at invadopodia. Results: We show here, for the first time, how RhoC activation is controlled at invadopodia and how this activation regulates cofilin phosphorylation to control cofilin's generation of actin-free barbed ends. Live-cell imaging of fluorescent RhoC biosensor reveals that RhoC activity is spatially confined to areas surrounding invadopodia. This spatiotemporal restriction of RhoC activity is controlled by "spatially distinct regulatory elements" that confine RhoC activation within this compartment. p190RhoGEF localizes around invadopodia to activate RhoC, whereas p190RhoGAP localizes inside invadopodia to deactivate the GTPase within the structure. RhoC activation enhances cofilin phosphorylation outside invadopodia. Conclusion: These results show how RhoC activity is spatially regulated at invadopodia by p190RhoGEF and p190RhoGAP. RhoC activation in areas surrounding invadopodia restricts cofilin activity to within the invadopodium core, resulting in a focused invadopodial protrusion. This mechanism likely enhances tumor cell invasion during metastasis.
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U2 - 10.1016/j.cub.2011.03.039
DO - 10.1016/j.cub.2011.03.039
M3 - Article
C2 - 21474314
AN - SCOPUS:79955425392
VL - 21
SP - 635
EP - 644
JO - Current Biology
JF - Current Biology
SN - 0960-9822
IS - 8
ER -