Yes-associated protein and p53-binding protein-2 interact through their WW and SH3 domains.
To understand the role of the Yes-associated protein (YAP), binding partners of its WW1 domain were isolated by a yeast two-hybrid screen. One of the interacting proteins was identified as p53-binding protein-2 (p53BP-2). YAP and p53BP-2 interacted in vitro and in vivo using their WW1 and SH3 domains, respectively. The ... YAP WW1 domain bound to the YPPPPY motif of p53BP-2, whereas the p53BP-2 SH3 domain interacted with the VPMRLR sequence of YAP, which is different from other known SH3 domain-binding motifs. By mutagenesis, we showed that this unusual SH3 domain interaction was due to the presence of three consecutive tryptophans located within the betaC strand of the SH3 domain. A point mutation within this triplet, W976R, restored the binding selectivity to the general consensus sequence for SH3 domains, the PXXP motif. A constitutively active form of c-Yes was observed to decrease the binding affinity between YAP and p53BP-2 using chloramphenicol acetyltransferase/enzyme-linked immunosorbent assay, whereas the overexpression of c-Yes did not modify this interaction. Since overexpression of an activated form of c-Yes resulted in tyrosine phosphorylation of p53BP-2, we propose that the p53BP-2 phosphorylation, possibly in the WW1 domain-binding motif, might negatively regulate the YAP.p53BP-2 complex.
Mesh Terms:
Adaptor Proteins, Signal Transducing, Amino Acid Motifs, Amino Acid Sequence, Apoptosis Regulatory Proteins, Blotting, Western, Brain, Carrier Proteins, Cell Line, Chloramphenicol O-Acetyltransferase, DNA, Complementary, Enzyme-Linked Immunosorbent Assay, Gene Library, Glutathione Transferase, Humans, Molecular Sequence Data, Mutagenesis, Site-Directed, Phosphoproteins, Phosphorylation, Plasmids, Point Mutation, Precipitin Tests, Protein Binding, Protein Structure, Tertiary, Sequence Homology, Amino Acid, Transfection, Tryptophan, Two-Hybrid System Techniques, Tyrosine, src Homology Domains
Adaptor Proteins, Signal Transducing, Amino Acid Motifs, Amino Acid Sequence, Apoptosis Regulatory Proteins, Blotting, Western, Brain, Carrier Proteins, Cell Line, Chloramphenicol O-Acetyltransferase, DNA, Complementary, Enzyme-Linked Immunosorbent Assay, Gene Library, Glutathione Transferase, Humans, Molecular Sequence Data, Mutagenesis, Site-Directed, Phosphoproteins, Phosphorylation, Plasmids, Point Mutation, Precipitin Tests, Protein Binding, Protein Structure, Tertiary, Sequence Homology, Amino Acid, Transfection, Tryptophan, Two-Hybrid System Techniques, Tyrosine, src Homology Domains
J. Biol. Chem.
Date: Apr. 27, 2001
PubMed ID: 11278422
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