title: A possible effector role for the pleckstrin homology (PH) domain of dynamin.
abstract: The large GTPase dynamin plays a key role in clathrin-mediated endocytosis in animal cells, although its mechanism of action remains unclear. Dynamins 1, 2, and 3 contain a pleckstrin homology (PH) domain that binds phosphoinositides with a very low affinity (K(D) > 1 mM), and this interaction appears to be crucial for function. These observations prompted the suggestion that an array of PH domains drives multivalent binding of dynamin oligomers to phosphoinositide-containing membranes. Although in vitro experiments reported here are consistent with this hypothesis, we find that PH domain mutations that abolish dynamin function do not alter localization of the protein in transfected cells, indicating that the PH domain does not play a simple targeting role. An alternative possibility is suggested by the geometry of dynamin helices resolved by electron microscopy. Even with one phosphatidylinositol-4,5-bisphosphate [PtdIns(4,5)P(2)] molecule bound per PH domain, these dynamin assemblies will elevate the concentration of PtdIns(4,5)P(2) at coated pit necks, and effectively cluster (or sequester) this phosphoinositide. In vitro fluorescence quenching studies using labeled phosphoinositides are consistent with dynamin-induced PtdIns(4,5)P(2) clustering. We therefore propose that the ability of dynamin to alter the local distribution of PtdIns(4,5)P(2) could be crucial for the role of this GTPase in promoting membrane scission during clathrin-mediated endocytosis. PtdIns(4,5)P(2) clustering could promote vesicle scission through direct effects on membrane properties, or might play a role in dynamin's ability to regulate actin polymerization.
authors: Department of Biochemistry and Biophysics, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-6059, USA.
authors: Kelley A
authors: KA
authors: Bethoney
authors: Megan C
authors: MC
authors: King
authors: Jenny E
authors: JE
authors: Hinshaw
authors: E Michael
authors: EM
authors: Ostap
authors: Mark A
authors: MA
authors: Lemmon
mesh_terms: D034281:Dynamins / Q000737:chemistry* / Q000378:metabolism; D004705:Endocytosis; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D017386:Sequence Homology, Amino Acid; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D019269:Phosphatidylinositol 4,5-Diphosphate; D034281:Dynamins