TC10 on endosomes regulates the local balance between microtubule stability and dynamics through the PAK2-JNK pathway and promotes axon outgrowth

The neuronal cytoskeleton comprises microtubules, actin filaments and neurofilaments, and plays a crucial role in axon outgrowth and transport. Microtubules and actin filaments have attracted considerable attention in axon regeneration studies. We have previously shown that TC10 (also known as RhoQ), a Rho family GTPase that promotes axon outgrowth through membrane addition, is required for efficient axon regeneration. This study demonstrates that TC10 on recycling endosomes, but not on the plasma membrane, balances microtubule stability and dynamics in the axons, thereby counteracting axon retraction. TC10 ablation reduced the phosphorylation of SCG10 (also known as STMN2) and MAP1B, which are neuronal microtubule-binding proteins and JNK substrates. Consistent with this, JNK phosphorylation was decreased in TC10-knockout neurons compared to in wild-type neurons. Furthermore, TC10 deletion significantly reduced PAK2 autophosphorylation. PAK2 was found on Rab11-positive endosomes in cell bodies and axons, and its localization to endosomes was reduced by TC10 loss. PAK inhibition reduced tubulin acetylation and JNK phosphorylation in axons. Furthermore, MKK4 and MKK7 (also known as MAP2K4 and MAP2K7, respectively) were found to mediate signaling from TC10-activated PAK to JNK on JIP1-positive endosomes. Overall, TC10 transmits a microtubule-regulatory signal from PAK2 to SCG10 and MAP1B via JNK on axonal endosomes.