Profilin binding couples chloride intracellular channel protein CLIC4 to RhoA-mDia2 signaling and filopodium formation

J Biol Chem. 2018 Dec 14;293(50):19161-19176. doi: 10.1074/jbc.RA118.002779. Epub 2018 Oct 31.


Chloride intracellular channel 4 (CLIC4) is a cytosolic protein implicated in diverse actin-based processes, including integrin trafficking, cell adhesion, and tubulogenesis. CLIC4 is rapidly recruited to the plasma membrane by RhoA-activating agonists and then partly colocalizes with β1 integrins. Agonist-induced CLIC4 translocation depends on actin polymerization and requires conserved residues that make up a putative binding groove. However, the mechanism and significance of CLIC4 trafficking have been elusive. Here, we show that RhoA activation by either lysophosphatidic acid (LPA) or epidermal growth factor is necessary and sufficient for CLIC4 translocation to the plasma membrane and involves regulation by the RhoA effector mDia2, a driver of actin polymerization and filopodium formation. We found that CLIC4 binds the G-actin-binding protein profilin-1 via the same residues that are required for CLIC4 trafficking. Consistently, shRNA-induced profilin-1 silencing impaired agonist-induced CLIC4 trafficking and the formation of mDia2-dependent filopodia. Conversely, CLIC4 knockdown increased filopodium formation in an integrin-dependent manner, a phenotype rescued by wild-type CLIC4 but not by the trafficking-incompetent mutant CLIC4(C35A). Furthermore, CLIC4 accelerated LPA-induced filopodium retraction. We conclude that through profilin-1 binding, CLIC4 functions in a RhoA-mDia2-regulated signaling network to integrate cortical actin assembly and membrane protrusion. We propose that agonist-induced CLIC4 translocation provides a feedback mechanism that counteracts formin-driven filopodium formation.

Keywords: G protein-coupled receptor; G-actin; Rho (Rho GTPase); actin; cell adhesion; cell motility; epidermal growth factor (EGF); filopodia; formin; lysophosphatidic acid; membrane trafficking; profilin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carrier Proteins / metabolism*
  • Cell Membrane / metabolism
  • Chloride Channels / chemistry
  • Chloride Channels / metabolism*
  • Chlorides / metabolism*
  • Conserved Sequence
  • Crystallography, X-Ray
  • Enzyme Activation
  • Formins
  • HeLa Cells
  • Humans
  • Integrins / metabolism
  • Models, Molecular
  • Profilins / chemistry
  • Profilins / metabolism*
  • Protein Binding
  • Protein Conformation
  • Protein Transport
  • Pseudopodia / metabolism*
  • Signal Transduction*
  • rhoA GTP-Binding Protein / metabolism*


  • CLIC4 protein, human
  • Carrier Proteins
  • Chloride Channels
  • Chlorides
  • DIAPH2 protein, human
  • Formins
  • Integrins
  • Profilins
  • rhoA GTP-Binding Protein

Associated data

  • PDB/2PAV
  • PDB/2AHE