c-Jun N-Terminal Kinase (JNK)-Interacting Protein-1b/Islet-Brain-1 Scaffolds Alzheimer's Amyloid Precursor Protein with JNK

• Published in: The Journal of neuroscience Vol. 21; no. 17; pp. 6597 - 6607
• Main Authors: Matsuda, Shuji, Yasukawa, Takashi, Homma, Yasuko, Ito, Yuko, Niikura, Takako, Hiraki, Takako, Hirai, Shuichi, Ohno, Shigeo, Kita, Yoshiko, Kawasumi, Masaoki, Kouyama, Keisuke, Yamamoto, Tokuo, Kyriakis, John M, Nishimoto, Ikuo
• Format: Journal Article
• Language: English
• Published: United States Soc Neuroscience 01.09.2001; Society for Neuroscience
• Subjects: Adaptor Proteins, Signal Transducing; Alzheimer Disease - metabolism; Amino Acid Motifs - physiology; Amino Acid Substitution; Amyloid beta-Protein Precursor - metabolism; Animals; Brain - metabolism; Carrier Proteins - genetics; Carrier Proteins - metabolism; Gene Library; Humans; islet-brain-1; JNK Mitogen-Activated Protein Kinases; JNK-interacting protein 1b; Mice; Mice, Inbred ICR; Mitogen-Activated Protein Kinases - metabolism; Mutagenesis, Site-Directed; Nerve Tissue Proteins - metabolism; Nuclear Proteins - genetics; Nuclear Proteins - metabolism; Phosphotyrosine - metabolism; Protein Binding - physiology; Protein Isoforms - genetics; Protein Isoforms - metabolism; Protein Structure, Tertiary - genetics; Protein Structure, Tertiary - physiology; Reverse Transcriptase Polymerase Chain Reaction; Structure-Activity Relationship; Trans-Activators - genetics; Trans-Activators - metabolism; Two-Hybrid System Techniques
• ISSN: 0270-6474; 1529-2401
• DOI: 10.1523/jneurosci.21-17-06597.2001

Using a yeast two-hybrid method, we searched for amyloid precursor protein (APP)-interacting molecules by screening mouse and human brain libraries. In addition to known interacting proteins containing a phosphotyrosine-interaction-domain (PID)-Fe65, Fe65L, Fe65L2, X11, and mDab1, we identified, as a novel APP-interacting molecule, a PID-containing isoform of mouse JNK-interacting protein-1 (JIP-1b) and its human homolog IB1, the established scaffold proteins for JNK. The APP amino acids Tyr(682), Asn(684), and Tyr(687) in the G(681)YENPTY(687) region were all essential for APP/JIP-1b interaction, but neither Tyr(653) nor Thr(668) was necessary. APP-interacting ability was specific for this additional isoform containing PID and was shared by both human and mouse homologs. JIP-1b expressed by mammalian cells was efficiently precipitated by the cytoplasmic domain of APP in the extreme Gly(681)-Asn(695) domain-dependent manner. Reciprocally, both full-length wild-type and familial Alzheimer's disease mutant APPs were precipitated by PID-containing JIP constructs. Antibodies raised against the N and C termini of JIP-1b coprecipitated JIP-1b and wild-type or mutant APP in non-neuronal and neuronal cells. Moreover, human JNK1beta1 formed a complex with APP in a JIP-1b-dependent manner. Confocal microscopic examination demonstrated that APP and JIP-1b share similar subcellular localization in transfected cells. These data indicate that JIP-1b/IB1 scaffolds APP with JNK, providing a novel insight into the role of the JNK scaffold protein as an interface of APP with intracellular functional molecules.