Abstract
Mammalian Enabled (Mena) is a mammalian homologue of Drosophila Enabled (Ena), which genetically interacts with Drosophila Abl tyrosine kinase. The signaling pathway involving c-Abl and Mena (Ena) is not fully understood. To find molecules that participate in the c-Abl/Mena pathway, we searched for Mena-binding proteins using a yeast two-hybrid system. We identified Abl interactor 1 (Abi-1), which is known to interact with c-Abl, as a binding protein for Mena. Binding analysis revealed that the Ena/Vasp homology 1 domain of Mena and the polyproline structure of Abi-1 are necessary for the interaction. The interaction between Mena and Abi-1 was also observed in a mammalian expression system. Importantly, Abi-1 dramatically promoted c-Abl-mediated tyrosine phosphorylation of Mena but not other substrates such as c-Cbl. Mutational analysis demonstrated that the phosphorylation site of Mena is Tyr-296. Our results suggest that Abi-1 regulates c-Abl-mediated phosphorylation of Mena by interacting with both proteins.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Signal Transducing*
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Alternative Splicing
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Amino Acid Sequence
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Animals
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Binding Sites
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Carrier Proteins / metabolism*
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Carrier Proteins / physiology
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Cell Line
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Cytoskeletal Proteins*
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DNA Mutational Analysis
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DNA, Complementary / metabolism
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DNA-Binding Proteins / metabolism*
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Gene Library
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Glutathione Transferase / metabolism
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Homeodomain Proteins / metabolism
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Homeodomain Proteins / physiology*
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Humans
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Mice
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Microfilament Proteins
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Microscopy, Fluorescence
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Molecular Sequence Data
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Phosphorylation
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Protein Binding
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Protein Structure, Tertiary
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Proto-Oncogene Proteins c-abl / metabolism*
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Rats
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Recombinant Fusion Proteins / metabolism
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Sequence Homology, Amino Acid
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Signal Transduction
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Two-Hybrid System Techniques
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Tyrosine / chemistry*
Substances
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ABI1 protein, human
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Abi1 protein, mouse
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Adaptor Proteins, Signal Transducing
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Carrier Proteins
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Cytoskeletal Proteins
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DNA, Complementary
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DNA-Binding Proteins
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ENA-VASP proteins
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Enah protein, mouse
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Homeodomain Proteins
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Microfilament Proteins
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Recombinant Fusion Proteins
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Tyrosine
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Glutathione Transferase
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Proto-Oncogene Proteins c-abl