PIP5K-Ras bistability initiates plasma membrane symmetry breaking to regulate cell polarity and migration

Symmetry breaking is a fundamental process that underlies key cellular behaviors such as cell polarity and migration, but the mechanism - how a uniform plasma membrane spontaneously transitions to an asymmetric state - is still unknown. Here in this study, using a combination of Dictyostelium amoeba, multiple mammalian leukocytes, and human cancer cells and 3D organoid systems, we monitored the localization dynamics of RasGTP and PIP5K, dissected the effects of eliminating, conditionally increasing, or optogenetically manipulating membrane PIP5K levels, screened for key regulators of Ras activation, and tracked single-molecules of PIP5K. Our data converge on a core biochemical circuit involving mutually inhibitory interactions between PIP5K and RasGTP that is both necessary and sufficient for symmetry breaking. The process is initiated by stochastic, localized Ras activation coupled with a decrease in the lifetime of the association of PIP5K with the membrane. A resulting localized reduction in PI(4,5)P2 facilitates the recruitment of a RasGEF to the corresponding domain, amplifying RasGTP production through a positive feedback loop. Dissociated PIP5K relocates to other membrane regions, where it suppresses Ras activation. These events separate the membrane into distinct active and inactive zones, even when receptor inputs or cytoskeletal activities are absent. This same core biochemical circuit controls the spatial organization of downstream PI3K/Akt/Rac signaling and actin/actomyosin activities, which generate the localized protrusions to define polarity and migration mode. While many models have been forwarded to explain polarity or symmetry breaking, the PIP5K-Ras mutually inhibitory bistable circuit we present here is the first universal molecular mechanism to explain the initiation of asymmetry as well as subsequent polarity and migration.