A Trio-Rac1-PAK1 signaling axis drives invadopodia disassembly

Rho family GTPases control cell migration and participate in the regulation of cancer metastasis. Invadopodia, associated with invasive tumor cells, are crucial for cellular invasion and metastasis. To study Rac1 GTPase in invadopodia dynamics, we developed a genetically-encoded, single-chain Rac1 Fluorescence Resonance Energy Transfer (FRET) biosensor. The biosensor shows Rac1 activity exclusion from the core of invadopodia, and higher activity when invadopodia disappear, suggesting that reduced Rac1 activity is necessary for their stability, and Rac1 activation is involved in disassembly. Photoactivating Rac1 at invadopodia confirmed this previously-unknown Rac1 function. We built an invadopodia disassembly model, where a signaling axis involving TrioGEF, Rac1, PAK1, and phosphorylation of cortactin, causing invadopodia dissolution. This mechanism is critical for the proper turnover of invasive structures during tumor cell invasion, where a balance of proteolytic activity and locomotory protrusions must be carefully coordinated to achieve a maximally invasive phenotype.