Abstract
Plectin is a major component of the cytoskeleton and is expressed in a wide variety of cell types. It plays an important role in the integrity of the cytoskeleton by cross-linking the three filamentous networks and stabilizing cell-matrix and cell-cell contacts. Sequence analysis showed that plectin contains a highly conserved actin-binding domain, consisting of a pair of calponin-like subdomains. Using yeast two-hybrid assays in combination with in vitro binding experiments, we demonstrate that the actin-binding domain of plectin is fully functional and preferentially binds to polymeric actin. The sequences required for actin binding were identified at the C-terminal end of the first calponin homology domain within the actin-binding domain of plectin. We found that the actin-binding domain of plectin is able to bundle actin filaments and we present evidence that this is mediated by the dimerization of this domain. In addition we also show that plectin and another member of the plakin family, dystonin, can heterodimerize by their actin-binding domains. We propose a new mechanism by which plectin and possibly also other actin-binding proteins can regulate the organization of the F-actin network in the cell.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Actins / chemistry
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Actins / genetics
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Actins / metabolism*
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Actins / ultrastructure
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Amino Acid Sequence
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Animals
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COS Cells
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Carrier Proteins*
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Cytoskeletal Proteins / chemistry
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Cytoskeletal Proteins / genetics
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Cytoskeletal Proteins / metabolism
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Cytoskeleton / chemistry
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Cytoskeleton / metabolism
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Cytoskeleton / ultrastructure
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Dimerization
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Dystonin
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Dystrophin / chemistry
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Dystrophin / genetics
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Dystrophin / metabolism
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Exons / genetics
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Fibroblasts
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Intermediate Filament Proteins / chemistry*
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Intermediate Filament Proteins / genetics
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Intermediate Filament Proteins / metabolism*
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Intermediate Filament Proteins / ultrastructure
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Kinetics
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Microscopy, Confocal
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Microscopy, Electron
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Molecular Sequence Data
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Nerve Tissue Proteins / chemistry
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Nerve Tissue Proteins / genetics
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Nerve Tissue Proteins / metabolism
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Peptide Fragments / chemistry
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Peptide Fragments / genetics
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Peptide Fragments / metabolism
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Peptide Fragments / ultrastructure
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Plectin
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Protein Binding
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Protein Isoforms / chemistry
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Protein Isoforms / genetics
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Protein Isoforms / metabolism
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Protein Isoforms / ultrastructure
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Protein Structure, Quaternary
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Protein Structure, Tertiary
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Rats
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Sequence Alignment
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Transfection
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Two-Hybrid System Techniques
Substances
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Actins
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Carrier Proteins
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Cytoskeletal Proteins
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Dst protein, mouse
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Dystonin
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Dystrophin
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Intermediate Filament Proteins
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Nerve Tissue Proteins
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Peptide Fragments
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Plec protein, rat
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Plectin
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Protein Isoforms