title: Phospholipids regulate localization and activity of mDia1 formin.
abstract: Diaphanous-related formins (DRFs) are large multi-domain proteins that nucleate and assemble linear actin filaments. Binding of active Rho family proteins to the GTPase-binding domain (GBD) triggers localization at the membrane and the activation of most formins if not all. In recent years GTPase regulation of formins has been extensively studied, but other molecular mechanisms that determine subcellular distribution or regulate formin activity have remained poorly understood. Here, we provide evidence that the activity and localization of mouse formin mDia1 can be regulated through interactions with phospholipids. The phospholipid-binding sites of mDia1 are clusters of positively charged residues in the N-terminal basic domain (BD) and at the C-terminal region. Upon binding to the lipid bilayer the N-terminal region of mDia1 induces strong clustering of phosphatidylinositol-4,5-bisphosphate (PIP(2)) and subsequently inserts into the membrane bilayer thus anchoring mDia1 to the reconstituted plasma membrane. In addition, an interaction of phospholipids with the C-terminal region of mDia1 causes a drastic reduction of its actin filament assembly activity. Our data suggest that the N-terminal phospholipid-binding sites help to anchor formins at the plasma membrane, and the interaction with phospholipids in the C-terminus functions as a switch for transient inactivation.
authors.affiliation: Institute for Anatomy and Cell Biology, Ludwig-Maximilians-Universitaet, Muenchen, Schillerstr. 42, 80336 Muenchen, Germany.
authors.forename: Nagendran
authors.initials: N
authors.lastname: Ramalingam
authors.forename: Hongxia
authors.initials: H
authors.lastname: Zhao
authors.forename: Dennis
authors.initials: D
authors.lastname: Breitsprecher
authors.forename: Pekka
authors.initials: P
authors.lastname: Lappalainen
authors.forename: Jan
authors.initials: J
authors.lastname: Faix
authors.forename: Michael
authors.initials: M
authors.lastname: Schleicher
mesh_terms: D008841:Actin Cytoskeleton / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites / Q000502:physiology; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D004789:Enzyme Activation / Q000502:physiology; D005326:Fetal Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D000080865:Formins; D008563:Membrane Lipids / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000502:physiology*; D051379:Mice; D008840:Microfilament Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D041681:NIH 3T3 Cells; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D010446:Peptide Fragments / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D010743:Phospholipids / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000502:physiology*; D058105:Polymerization; D011485:Protein Binding / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology
chemical_list: D002352:Carrier Proteins; C491464:Diap1 protein, mouse; D005326:Fetal Proteins; D000080865:Formins; D008563:Membrane Lipids; D008840:Microfilament Proteins; D009687:Nuclear Proteins; D010446:Peptide Fragments; D010743:Phospholipids