Zyxin interacts with the SH3 domains of the cytoskeletal proteins LIM-nebulette and Lasp-1.
Zyxin is a versatile component of focal adhesions in eukaryotic cells. Here we describe a novel binding partner of zyxin, which we have named LIM-nebulette. LIM-nebulette is an alternative splice variant of the sarcomeric protein nebulette, which, in contrast to nebulette, is expressed in non-muscle cells. It displays a modular ... structure with an N-terminal LIM domain, three nebulin-like repeats, and a C-terminal SH3 domain and shows high similarity to another cytoskeletal protein, Lasp-1 (LIM and SH3 protein-1). Co-precipitation studies and results obtained with the two-hybrid system demonstrate that LIM-nebulette and Lasp-1 interact specifically with zyxin. Moreover, the SH3 domain from LIM-nebulette is both necessary and sufficient for zyxin binding. The SH3 domains from Lasp-1 and nebulin can also interact with zyxin, but the SH3 domains from more distantly related proteins such as vinexin and sorting nexin 9 do not. On the other hand, the binding site in zyxin is situated at the extreme N terminus as shown by site-directed mutagenesis. LIM-nebulette and Lasp-1 use the same linear binding motif. This motif shows some similarity to a class II binding site but does not contain the classical PXXP sequence. LIM-nebulette reveals a subcellular distribution at focal adhesions similar to Lasp-1. Thus, LIM-nebulette, Lasp-1, and zyxin may play an important role in the organization of focal adhesions.
Mesh Terms:
Adaptor Proteins, Signal Transducing, Alternative Splicing, Amino Acid Motifs, Amino Acid Sequence, Animals, Binding Sites, Blotting, Northern, Carrier Proteins, Cloning, Molecular, Cytoskeletal Proteins, Cytoskeleton, DNA, Complementary, Exons, Focal Adhesions, Glutathione Transferase, Glycoproteins, Green Fluorescent Proteins, Hela Cells, Homeodomain Proteins, Humans, Luminescent Proteins, Mice, Microscopy, Fluorescence, Molecular Sequence Data, Muscle Proteins, Mutagenesis, Site-Directed, Neoplasm Proteins, Plasmids, Protein Binding, Protein Structure, Tertiary, Sequence Homology, Amino Acid, Tissue Distribution, Transfection, Two-Hybrid System Techniques, Vesicular Transport Proteins, src Homology Domains
Adaptor Proteins, Signal Transducing, Alternative Splicing, Amino Acid Motifs, Amino Acid Sequence, Animals, Binding Sites, Blotting, Northern, Carrier Proteins, Cloning, Molecular, Cytoskeletal Proteins, Cytoskeleton, DNA, Complementary, Exons, Focal Adhesions, Glutathione Transferase, Glycoproteins, Green Fluorescent Proteins, Hela Cells, Homeodomain Proteins, Humans, Luminescent Proteins, Mice, Microscopy, Fluorescence, Molecular Sequence Data, Muscle Proteins, Mutagenesis, Site-Directed, Neoplasm Proteins, Plasmids, Protein Binding, Protein Structure, Tertiary, Sequence Homology, Amino Acid, Tissue Distribution, Transfection, Two-Hybrid System Techniques, Vesicular Transport Proteins, src Homology Domains
J. Biol. Chem.
Date: May. 07, 2004
PubMed ID: 15004028
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