title: Reversible chemical dimerizer-induced recovery of PIP2 levels moves clathrin to the plasma membrane.
abstract: Chemical dimerizers are powerful non-invasive tools for bringing molecules together inside intact cells. We recently introduced a rapidly reversible chemical dimerizer system which enables transient translocation of enzymes to and from the plasma membrane (PM). Here we have applied this system to transiently activate phosphatidylinositol 4,5-bisphosphate (PIP2) breakdown at the PM via translocation of phosphoinositide 5-phosphatase (5Ptase). We found that the PIP2 sensor phospholipase C-δ PH domain (PLCδ-PH) is released from the PM upon addition of the reversible chemical dimerizer rCD1. By outcompeting rCD1, rapid release of the 5Ptase from the PM is followed by PIP2 recovery. This permits the observation of the PIP2-dependent clathrin assembly at the PM.
authors.affiliation: European Molecular Biology Laboratory (EMBL), Meyerhofstr. 1, 69117 Heidelberg, Germany.
authors.forename: Martina
authors.initials: M
authors.lastname: Schifferer
authors.affiliation: European Molecular Biology Laboratory (EMBL), Meyerhofstr. 1, 69117 Heidelberg, Germany.
authors.forename: Suihan
authors.initials: S
authors.lastname: Feng
authors.affiliation: European Molecular Biology Laboratory (EMBL), Meyerhofstr. 1, 69117 Heidelberg, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Stein
authors.affiliation: European Molecular Biology Laboratory (EMBL), Meyerhofstr. 1, 69117 Heidelberg, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Tischer
authors.affiliation: European Molecular Biology Laboratory (EMBL), Meyerhofstr. 1, 69117 Heidelberg, Germany. Electronic address: schultz@embl.de.
authors.forename: Carsten
authors.initials: C
authors.lastname: Schultz
mesh_terms: D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D002966:Clathrin / Q000378:metabolism*; D019281:Dimerization; D004705:Endocytosis / Q000187:drug effects; D006801:Humans; D010716:Phosphatidylinositols / Q000737:chemistry / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects*; D000099182:Phosphoinositide 5-Phosphatases
chemical_list: D002966:Clathrin; D010716:Phosphatidylinositols; D010744:Phosphoric Monoester Hydrolases; D000099182:Phosphoinositide 5-Phosphatases
keywords: Adapter proteins; Clathrin; Endocytosis; Phosphatidylinositol 4,5-bisphosphate; Phosphoinositol 5-phosphatase