TY - JOUR
T1 - Tks5 recruits AFAP-110, p190RhoGAP, and cortactin for podosome formation
AU - Crimaldi, Luca
AU - Courtneidge, Sara A.
AU - Gimona, Mario
N1 - Funding Information:
The Gimona laboratory is supported by a Marie Curie Excellence Grant (MEXT-CT-2003-002573) of the European Union. L.C. is grateful to Dr. Alberto Luini (Consorzio Mario Negri Sud, Italy) for providing support throughout this work. Support from the National Cancer Institute (to S.A.C.) is gratefully acknowledged.
Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 2009/9/10
Y1 - 2009/9/10
N2 - Podosome formation in vascular smooth muscle cells is characterized by the recruitment of AFAP-110, p190RhoGAP, and cortactin, which have specific roles in Src activation, local down-regulation of RhoA activity, and actin polymerization, respectively. However, the molecular mechanism that underlies their specific recruitment to podosomes remains unknown. The scaffold protein Tks5 is localized to podosomes in Src-transformed fibroblasts and in smooth muscle cells, and may serve as a specific recruiting adapter for various components during podosome formation. We show here that induced mislocalization of Tks5 to the surface of mitochondria leads to a major subcellular redistribution of AFAP-110, p190RhoGAP, and cortactin, and to inhibition of podosome formation. Analysis of a series of similarly mistargeted deletion mutants of Tks5 indicates that the fifth SH3 domain is essential for this recruitment. A Tks5 mutant lacking the PX domain also inhibits podosome formation and induces the redistribution of AFAP-110, p190RhoGAP, and cortactin to the perinuclear area. By expressing a catalytically inactive point mutant and by siRNA-mediated expression knock-down we also provide evidence that p190RhoGAP is required for podosome formation. Together our findings demonstrate that Tks5 plays a central role in the recruitment of AFAP-110, p190RhoGAP, and cortactin to drive podosome formation.
AB - Podosome formation in vascular smooth muscle cells is characterized by the recruitment of AFAP-110, p190RhoGAP, and cortactin, which have specific roles in Src activation, local down-regulation of RhoA activity, and actin polymerization, respectively. However, the molecular mechanism that underlies their specific recruitment to podosomes remains unknown. The scaffold protein Tks5 is localized to podosomes in Src-transformed fibroblasts and in smooth muscle cells, and may serve as a specific recruiting adapter for various components during podosome formation. We show here that induced mislocalization of Tks5 to the surface of mitochondria leads to a major subcellular redistribution of AFAP-110, p190RhoGAP, and cortactin, and to inhibition of podosome formation. Analysis of a series of similarly mistargeted deletion mutants of Tks5 indicates that the fifth SH3 domain is essential for this recruitment. A Tks5 mutant lacking the PX domain also inhibits podosome formation and induces the redistribution of AFAP-110, p190RhoGAP, and cortactin to the perinuclear area. By expressing a catalytically inactive point mutant and by siRNA-mediated expression knock-down we also provide evidence that p190RhoGAP is required for podosome formation. Together our findings demonstrate that Tks5 plays a central role in the recruitment of AFAP-110, p190RhoGAP, and cortactin to drive podosome formation.
KW - AFAP-110
KW - Cortactin
KW - Podosomes
KW - Smooth muscle cells
KW - Tks5
KW - p190RhoGAP
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U2 - 10.1016/j.yexcr.2009.06.012
DO - 10.1016/j.yexcr.2009.06.012
M3 - Article
C2 - 19540230
AN - SCOPUS:67651161854
VL - 315
SP - 2581
EP - 2592
JO - Experimental Cell Research
JF - Experimental Cell Research
SN - 0014-4827
IS - 15
ER -