α-Synuclein facilitates endocytosis by elevating the steady-state levels of phosphatidylinositol 4,5-bisphosphate

-Synuclein (-Syn) is a protein implicated in the pathogenesis of Parkinsons disease (PD). It is an intrinsically disordered protein that binds acidic phospholipids. Growing evidence supports a role for -Syn in membrane trafficking, including, mechanisms of endocytosis and exocytosis, although the exact role of -Syn in these mechanisms is currently unclear. Here we have investigated the role of -Syn in membrane trafficking through its association with acidic phosphoinositides (PIPs), such as phosphatidylinositol 4,5-bisphosphate (PI4,5P2) and phosphatidylinositol 3,4-bisphosphate (PI3,4P2). Our results show that -Syn colocalizes with PIP2 and the phosphorylated active form of the clathrin adaptor AP2 at clathrin-coated pits. Using endocytosis of transferrin, an indicator of clathrin mediated endocytosis (CME), we find that -Syn involvement in endocytosis is specifically mediated through PI4,5P2 levels. We further show that the rate of synaptic vesicle (SV) endocytosis is differentially affected by -Syn mutations. In accord with their effects on PI4,5P2 levels at the plasma membrane, the PD associated E46K and A53T mutations further enhance SV endocytosis. However, neither A30P mutation, nor Lysine to Glutamic acid substitutions at the KTKEGV repeat domain of -Syn, that interfere with phospholipid binding, affect SV endocytosis. This study provides evidence for a critical involvement of PIPs in -Syn-mediated membrane trafficking. Significance Statement-Synuclein (-Syn) protein is known for its causative role in Parkinsons disease. -Syn is normally involved in mechanisms of membrane trafficking, including endocytosis, exocytosis and synaptic vesicles cycling. However, a certain degree of controversy regarding the exact role of -Syn in these mechanisms persists. Here we show that -Syn acts to increase plasma membrane levels PI4,5P2 and PI3,4P2 to facilitate clathrin mediated and synaptic vesicles endocytosis. Based on the results, we suggest that -Syn interactions with the acidic phosphoinositides facilitate a shift in their homeostasis to support endocytosis.