Phosphatidylinositol 4,5-bisphosphate (PIP2) has been shown involved in key chemotactic signaling pathways in Dictyostelium, but our understanding of its roles in this signaling network is very limited. To explore the hypothesis that PIP2 negatively regulate chemotactic signaling events, we used the Chemically-induced Dimerization system, which allows inducible translocation of cellular proteins, to synthetically manipulate PIP2 level and dissect the signaling events. In my current data, after I synthetically recruited the PIP2-specific 5-phosphotase Inp54p to the plasma membrane, the PIP2 biosensor PHplcdelta fell off the membrane and the cells robustly oscillated between a spreading and a crunching morphology. Based on the localization changes of PIP2-binding protein PTEN, the PIP2 levels may further go down as cells spread, and rebound as cells crunch. In the same time, cells with the spreading morphology, but not the other one, have clearly activated chemotactic signaling events as shown by Ras and PIP3 biosensors, as well as highly elevated F-actin along the periphery. Further, cells still carried out this response when PIP3 production was diminished by the PI 3-Kinase inhibitor LY. Remarkably, these signaling events seemed to prevail when actin polymerization was greatly inhibited, as the signaling molecule PTEN showed a distinct spiral wave pattern in LatA-treated cells. These suggest PIP2 negatively regulates the chemotactic signaling network, at least partially independent of actin. We propose that PIP2 serves as an inhibitory role on the upstream chemotactic signaling molecule Ras; when PIP2 levels are decreased by Inp54p recruitment, multiple parallel chemotactic signaling pathways get activated as a result of Ras activation. Also in combine with previous evidence, we propose that PI 5-Kinase activation is downstream of PKBs activation, which up-regulates PIP2 levels, shuts down the signaling events and leads to the crunching morphology following spreading.