title: The IQGAP1 protein is a calmodulin-regulated barbed end capper of actin filaments: possible implications in its function in cell migration.
abstract: IQGAP1 is a large modular protein that displays multiple partnership and is thought to act as a scaffold in coupling cell signaling to the actin and microtubule cytoskeletons in cell migration, adhesion, and cytokinesis. However the molecular mechanisms underlying the activities of IQGAP1 are poorly understood in part because of its large size, poor solubility and lack of functional assays to challenge biochemical properties in various contexts. We have purified bacterially expressed recombinant human IQGAP1. The protein binds Cdc42, Rac1, and the CRIB domain of N-WASP in a calmodulin-sensitive fashion. We further show that in addition to bundling of filaments via a single N-terminal calponin-homology domain, IQGAP1 actually regulates actin assembly. It caps barbed ends, with a higher affinity for ADP-bound terminal subunits (K(B) = 4 nM). The barbed end capping activity is inhibited by calmodulin, consistent with calmodulin binding to IQGAP1 with a K(C) of 40 nm, both in the absence and presence of Ca(2+) ions. The barbed end capping activity resides in the C-terminal half of IQGAP1. It is possible that the capping activity of IQGAP1 accounts for its stimulation of cell migration. We further find that bacterially expressed recombinant IQGAP1 fragments easily co-purify with nucleic acids that turn out to activate N-WASP protein to branch filaments with Arp2/3 complex. The present results open perspectives for tackling the function of IQGAP1 in more complex reconstituted systems.
authors: Cytoskeleton Dynamics and Motility group, UPR 3289, Laboratoire d'Enzymologie et Biochimie Structurale, CNRS, 91198 Gif-sur-Yvette, France.
authors: Andrea
authors: A
authors: Pelikan-Conchaudron
authors: Christophe
authors: C
authors: Le Clainche
authors: Dominique
authors: D
authors: Didry
authors: Marie-France
authors: MF
authors: Carlier
mesh_terms: D000199:Actins / Q000378:metabolism; D000244:Adenosine Diphosphate / Q000737:chemistry; D002147:Calmodulin / Q000378:metabolism; D002465:Cell Movement; D018076:DNA, Complementary / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D017354:Point Mutation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D015398:Signal Transduction; D012995:Solubility; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal / Q000378:metabolism; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism; D020703:ras GTPase-Activating Proteins / Q000378:metabolism*
chemical_list: D000199:Actins; D002147:Calmodulin; D018076:DNA, Complementary; C101203:IQ motif containing GTPase activating protein 1; D011994:Recombinant Proteins; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal; D020703:ras GTPase-Activating Proteins; D000244:Adenosine Diphosphate; D020764:cdc42 GTP-Binding Protein