title: Integrating the actin and vimentin cytoskeletons. adhesion-dependent formation of fimbrin-vimentin complexes in macrophages.
abstract: Cells adhere to the substratum through specialized structures that are linked to the actin cytoskeleton. Recent studies report that adhesion also involves the intermediate filament (IF) and microtubule cytoskeletons, although their mechanisms of interaction are unknown. Here we report evidence for a novel adhesion-dependent interaction between components of the actin and IF cytoskeletons. In biochemical fractionation experiments, fimbrin and vimentin coprecipitate from detergent extracts of macrophages using vimentin- or fimbrin-specific antisera. Fluorescence microscopy confirms the biochemical association. Both proteins colocalized to podosomes in the earliest stages of cell adhesion and spreading. The complex is also found in filopodia and retraction fibers. After detergent extraction, fimbrin and vimentin staining of podosomes, filopodia, and retraction fibers are lost, confirming that the complex is localized to these structures. A 1:4 stoichiometry of fimbrin binding to vimentin and a low percentage (1%) of the extracted vimentin suggest that fimbrin interacts with a vimentin subunit. A fimbrin-binding site was identified in the NH(2)-terminal domain of vimentin and the vimentin binding site at residues 143-188 in the CH1 domain of fimbrin. Based on these observations, we propose that a fimbrin-vimentin complex may be involved in directing the assembly of the vimentin cytoskeleton at cell adhesion sites.
authors.affiliation: Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02142, USA. correia@wi.mit.edu
authors.forename: I
authors.initials: I
authors.lastname: Correia
authors.forename: D
authors.initials: D
authors.lastname: Chu
authors.forename: Y H
authors.initials: YH
authors.lastname: Chou
authors.forename: R D
authors.initials: RD
authors.lastname: Goldman
authors.forename: P
authors.initials: P
authors.lastname: Matsudaira
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D002448:Cell Adhesion; D002460:Cell Line; D016764:Cell Polarity; D048429:Cell Size; D003599:Cytoskeleton / Q000378:metabolism*; D007382:Intermediate Filaments / Q000378:metabolism; D008264:Macrophages / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008840:Microfilament Proteins; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011108:Polymers; D011485:Protein Binding; D011554:Pseudopodia / Q000378:metabolism; D017384:Sequence Deletion; D014746:Vimentin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D000199:Actins; D008562:Membrane Glycoproteins; D008840:Microfilament Proteins; D010446:Peptide Fragments; D011108:Polymers; D014746:Vimentin; C026742:plastin