RhoJ integrates attractive and repulsive cues in directional migration of endothelial cells.

During angiogenesis,VEGFacts as an attractive cue for endothelial cells (ECs), while Sema3E mediates repulsive cues. Here, we show that the smallGTPase RhoJ integrates these opposing signals in directionalECmigration. In theGTP-bound state, RhoJ interacts with the cytoplasmic domain of PlexinD1. Upon Sema3E stimulation, RhoJ released from PlexinD1 induces cell contraction. PlexinD1-bound RhoJ further facilitates Sema3E-induced PlexinD1-VEGFR2 association,VEGFR2 transphosphorylation at Y1214, and p38MAPKactivation, leading to reverseECmigration. UponVEGFstimulation, RhoJ is required for the formation of the holoreceptor complex comprisingVEGFR2, PlexinD1, and neuropilin-1, thereby preventing degradation of internalizedVEGFR2, prolonging downstream signal transductions viaPLC gamma, Erk, and Akt, and promoting forwardECmigration. After conversion to theGDP-bound state, RhoJ shifts from PlexinD1 toVEGFR2, which then terminates theVEGFR2 signals. RhoJ deficiency inECs efficiently suppressed aberrant angiogenesis in ischemic retina. These findings suggest that distinct RhoGTPases may act as context-dependent integrators of chemotactic cues in directional cell migration and may serve as candidate therapeutic targets to manipulate cell motility in disease or tissue regeneration.