TY - JOUR
T1 - Tks5 and SHIP2 regulate invadopodium maturation, but not initiation, in breast carcinoma cells
AU - Sharma, Ved P.
AU - Eddy, Robert
AU - Entenberg, David
AU - Kai, Masayuki
AU - Gertler, Frank B.
AU - Condeelis, John
N1 - Funding Information:
We thank the investigators listed in the Supplemental Experimental Procedures for generous sharing of cells and reagents; Xiaoming Chen for the Tks5-R42A/R93A mutant; the Analytical Imaging Facility and Gruss Lipper Biophotonics Center at Albert Einstein College of Medicine for microscopy help; and members of the Condeelis, Segall, Cox, and Hodgson laboratories for guidance. This work was funded by NIH grants CA150344 and U54-CA112967 (F.B.G.) and a postdoctoral fellowship from Susan G. Komen for the Cure KG111405 to V.P.S.
PY - 2013/11/4
Y1 - 2013/11/4
N2 - Background: Tks5 regulates invadopodium formation, but the precise timing during invadopodium lifetime (initiation, stabilization, maturation) when Tks5 plays a role is not known. Results: We report new findings based on high-resolution spatiotemporal live-cell imaging of invadopodium precursor assembly. Cortactin, N-WASP, cofilin, and actin arrive together to form the invadopodium precursor, followed by Tks5 recruitment. Tks5 is not required for precursor initiation but is needed for precursor stabilization, which requires the interaction of the phox homology (PX) domain of Tks5 with PI(3,4)P 2. During precursor formation, PI(3,4)P2 is uniformly distributed but subsequently starts accumulating at the precursor core 3-4 min after core initiation, and conversely, PI(3,4,5)P3 gets enriched in a ring around the precursor core. SHIP2, a 5′-inositol phosphatase, localizes at the invadopodium core and regulates PI(3,4)P2 levels locally at the invadopodium. The timing of SHIP2 arrival at the invadopodium precursor coincides with the onset of PI(3,4)P2 accumulation. Consistent with its late arrival, we found that SHIP2 inhibition does not affect precursor formation but does cause decreases in mature invadopodia and matrix degradation, whereas SHIP2 overexpression increases matrix degradation. Conclusions: Together, these findings lead us to propose a new sequential model that provides novel insights into molecular mechanisms underlying invadopodium precursor initiation, stabilization, and maturation into a functional invadopodium.
AB - Background: Tks5 regulates invadopodium formation, but the precise timing during invadopodium lifetime (initiation, stabilization, maturation) when Tks5 plays a role is not known. Results: We report new findings based on high-resolution spatiotemporal live-cell imaging of invadopodium precursor assembly. Cortactin, N-WASP, cofilin, and actin arrive together to form the invadopodium precursor, followed by Tks5 recruitment. Tks5 is not required for precursor initiation but is needed for precursor stabilization, which requires the interaction of the phox homology (PX) domain of Tks5 with PI(3,4)P 2. During precursor formation, PI(3,4)P2 is uniformly distributed but subsequently starts accumulating at the precursor core 3-4 min after core initiation, and conversely, PI(3,4,5)P3 gets enriched in a ring around the precursor core. SHIP2, a 5′-inositol phosphatase, localizes at the invadopodium core and regulates PI(3,4)P2 levels locally at the invadopodium. The timing of SHIP2 arrival at the invadopodium precursor coincides with the onset of PI(3,4)P2 accumulation. Consistent with its late arrival, we found that SHIP2 inhibition does not affect precursor formation but does cause decreases in mature invadopodia and matrix degradation, whereas SHIP2 overexpression increases matrix degradation. Conclusions: Together, these findings lead us to propose a new sequential model that provides novel insights into molecular mechanisms underlying invadopodium precursor initiation, stabilization, and maturation into a functional invadopodium.
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U2 - 10.1016/j.cub.2013.08.044
DO - 10.1016/j.cub.2013.08.044
M3 - Article
C2 - 24206842
AN - SCOPUS:84887161574
SN - 0960-9822
VL - 23
SP - 2079
EP - 2089
JO - Current Biology
JF - Current Biology
IS - 21
ER -