Phospholipids regulate localization and activity of mDia1 formin

• Published in: European journal of cell biology 2010-10, Vol.89 (10), p.723-732
• Main Authors: Ramalingam, Nagendran, Zhao, Hongxia, Breitsprecher, Dennis, Lappalainen, Pekka, Faix, Jan, Schleicher, Michael
• Format: Article
• Language: English
• Subjects: Actin Cytoskeleton - chemistry; Actin Cytoskeleton - metabolism; Actin polymerization and phospholipids; Actin-binding proteins; Animals; Binding Sites - physiology; Carrier Proteins - antagonists & inhibitors; Carrier Proteins - chemistry; Carrier Proteins - metabolism; Cell Membrane - chemistry; Cell Membrane - metabolism; Enzyme Activation - physiology; Fetal Proteins - antagonists & inhibitors; Fetal Proteins - chemistry; Fetal Proteins - metabolism; Formins; mDia; Membrane Lipids - antagonists & inhibitors; Membrane Lipids - chemistry; Membrane Lipids - physiology; Mice; Microfilament Proteins - antagonists & inhibitors; Microfilament Proteins - chemistry; Microfilament Proteins - metabolism; NIH 3T3 Cells; Nuclear Proteins - antagonists & inhibitors; Nuclear Proteins - chemistry; Nuclear Proteins - metabolism; Peptide Fragments - antagonists & inhibitors; Peptide Fragments - chemistry; Peptide Fragments - metabolism; Phospholipids - antagonists & inhibitors; Phospholipids - chemistry; Phospholipids - physiology; Polymerization; Protein Binding - physiology; Protein Structure, Tertiary - physiology
• ISSN: 0171-9335; 1618-1298
• DOI: 10.1016/j.ejcb.2010.06.001

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.