Enforced expression of phosphatidylinositol 4-phosphate 5-kinase homolog (PIPKH) alters phosphatidylinositol 4,5-bisphosphate distribution and the localization of small G-proteins

Abstract

The generation of phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) by phosphatidylinositol 4-phosphate 5-kinases (PIP5Ks) is essential for many of the functions including the control of cytoskeleton, signal transduction and endocytosis. Additionally, due to its presence in the plasma membrane and its anionic charge PtdIns(4,5)P2, together with phosphatidylserine, imbue the inner leaflet of the plasma membrane with a negative surface charge. This negative charge helps to define the identity of the plasma membrane as serves to recruit or regulate a multitude of proteins that contain polybasic domains or patches. Here we determine that the phosphatidylinositol 4-phosphate 5-kinase homolog (PIPKH) alters the subcellular distribution of PtdIns(4,5)P2 by re-localizing the PIP5Ks to endomembranes. Consistently, we find a redistribution of the PIP5K family members to endomembrane structures upon PIPKH overexpression that is accompanied by an accumulation of PtdIns(4,5)P2 and phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P3), which further influences the distribution of endosomes and lysosomes. Additionally, we demonstrate that the accumulation of polyphosphoinositides increases their negative surface charge that in turn leads to the relocalization of surface charge probes as well as the polycationic proteins K-Ras and Rac1.