Phosphatidylinositol polyphosphate binding to the mammalian septin H5 is modulated by GTP.

Septins are members of a conserved family of GTPases found in organisms as diverse as budding yeast and mammals. In budding yeast, septins form hetero-oligomeric filaments that lie adjacent to the membrane at the mother-bud neck, whereas in mammals, they concentrate at the cleavage furrow of mitotic cells; in both cases, septins provide a required function for cytokinesis. What directs the location and determines the stability of septin filaments, however, remains unknown. Here we show that the mammalian septin H5 is associated with the plasma membrane and specifically binds the phospholipids phosphatidylinositol 4, 5-bisphosphate (PtdIns(4,5)P(2)) and phosphatidylinositol 3,4, 5-trisphosphate (PtdIns(3,4,5)P(3)). Deletion analysis revealed that this binding occurs at a site rich in basic residues that is conserved in most septins and is located adjacent to the GTP-binding motif. Phosphoinositide binding was inhibited by mutations within this motif and was also blocked by agents known to associate with PtdInsP(2) or by a peptide corresponding to the predicted PtdInsP(2)-binding sequence of H5. GTP binding and hydrolysis by H5 significantly reduced its PtdInsP(2)-binding capability. Treatment of cells with agents that occluded, dephosphorylated or degraded PtdInsP(2) altered the appearance and localization of H5. These results indicate that the interaction of septins with PtdInsP(2) might be an important cellular mechanism for the spatial and temporal control of septin accumulation.