title: The Dictyostelium CARMIL protein links capping protein and the Arp2/3 complex to type I myosins through their SH3 domains.
abstract: Fusion proteins containing the Src homology (SH)3 domains of Dictyostelium myosin IB (myoB) and IC (myoC) bind a 116-kD protein (p116), plus nine other proteins identified as the seven member Arp2/3 complex, and the alpha and beta subunits of capping protein. Immunoprecipitation reactions indicate that myoB and myoC form a complex with p116, Arp2/3, and capping protein in vivo, that the myosins bind to p116 through their SH3 domains, and that capping protein and the Arp2/3 complex in turn bind to p116. Cloning of p116 reveals a protein dominated by leucine-rich repeats and proline-rich sequences, and indicates that it is a homologue of Acan 125. Studies using p116 fusion proteins confirm the location of the myosin I SH3 domain binding site, implicate NH(2)-terminal sequences in binding capping protein, and show that a region containing a short sequence found in several G-actin binding proteins, as well as an acidic stretch, can activate Arp2/3-dependent actin nucleation. p116 localizes along with the Arp2/3 complex, myoB, and myoC in dynamic actin-rich cellular extensions, including the leading edge of cells undergoing chemotactic migration, and dorsal, cup-like, macropinocytic extensions. Cells lacking p116 exhibit a striking defect in the formation of these macropinocytic structures, a concomitant reduction in the rate of fluid phase pinocytosis, a significant decrease in the efficiency of chemotactic aggregation, and a decrease in cellular F-actin content. These results identify a complex that links key players in the nucleation and termination of actin filament assembly with a ubiquitous barbed end-directed motor, indicate that the protein responsible for the formation of this complex is physiologically important, and suggest that previously reported myosin I mutant phenotypes in Dictyostelium may be due, at least in part, to defects in the assembly state of actin. We propose that p116 and Acan 125, along with homologues identified in Caenorhabditis elegans, Drosophila, mouse, and man, be named CARMIL proteins, for capping protein, Arp2/3, and myosin I linker.
authors: Laboratory of Cell Biology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors: G
authors: G
authors: Jung
authors: K
authors: K
authors: Remmert
authors: X
authors: X
authors: Wu
authors: J M
authors: JM
authors: Volosky
authors: J A
authors: JA
authors: Hammer
mesh_terms: D051339:Actin Depolymerizing Factors; D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D000199:Actins / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites / Q000502:physiology; D017173:Caenorhabditis elegans; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D021961:Cell Membrane Structures / Q000502:physiology; D002465:Cell Movement / Q000502:physiology; D002633:Chemotaxis / Q000502:physiology; D003001:Cloning, Molecular; D003598:Cytoskeletal Proteins; D051345:Destrin; D004023:Dictyostelium; D004330:Drosophila; D004705:Endocytosis / Q000502:physiology; D006801:Humans; D046911:Macromolecular Substances; D051379:Mice; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009218:Myosins / Q000378:metabolism*; D011233:Precipitin Tests; D011485:Protein Binding / Q000502:physiology; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D018909:src Homology Domains / Q000502:physiology*
chemical_list: C495926:ACTR2 protein, human; C495928:ACTR3 protein, human; C096372:Acan125 protein, Acanthamoeba; D051339:Actin Depolymerizing Factors; D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D000199:Actins; C495929:Actr3 protein, mouse; C430925:CARMIL protein, Dictyostelium; D002352:Carrier Proteins; D003598:Cytoskeletal Proteins; D051345:Destrin; D046911:Macromolecular Substances; D008840:Microfilament Proteins; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; D009218:Myosins