title: NF-kappa B p65 transactivation domain is involved in the NF-kappa B-inducing kinase pathway.
abstract: NF-kappa B-inducing kinase (NIK) is involved in the signal transduction pathway leading to the NF-kappa B activation. In this report, we demonstrate that the NIK-mediated NF-kappa B activation involves the transactivation (TA) domain of p65 subunit of NF-kappa B and the nuclear translocation of IKK alpha. By using luciferase assay, we found that both IKK alpha and IKK beta could activate NF-kappa B in synergy with NIK. Interestingly, although IKK beta stimulated the NIK-mediated I kappa B degradation, IKK alpha stimulated the action of NF-kappa B without enhancing the I kappa B degradation. By using heterologous transactivation system with Gal4 DNA-binding domain in fusion with various portions of p65 TA domain, we found that the transactivation domain 1 (TA1) of p65 serves as the direct target for the NIK-IKK alpha cascade and that the serine residue at 536 within p65 TA1 is indispensable for this action. Furthermore, we found that this action of NIK depends on the energy-dependent action of Ras-related protein (Ran) since the dominant negative mutant of Ran (RanQ69L) inhibited the transcriptional activity of p65 by preventing the nuclear import of IKK alpha.
authors.affiliation: Department of Molecular Genetics, Nagoya City University Medical School, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya 467-8601, Japan.
authors.forename: Xu
authors.initials: X
authors.lastname: Jiang
authors.forename: Naoko
authors.initials: N
authors.lastname: Takahashi
authors.forename: Kiichiro
authors.initials: K
authors.lastname: Ando
authors.forename: Takanobu
authors.initials: T
authors.lastname: Otsuka
authors.forename: Toshifumi
authors.initials: T
authors.lastname: Tetsuka
authors.forename: Takashi
authors.initials: T
authors.lastname: Okamoto
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000502:physiology*; D000818:Animals; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006801:Humans; D051550:I-kappa B Kinase; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D051996:Transcription Factor RelA; D015533:Transcriptional Activation; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D020931:ran GTP-Binding Protein / Q000235:genetics / Q000378:metabolism; D000097782:NF-kappaB-Inducing Kinase
chemical_list: D004268:DNA-Binding Proteins; D051550:I-kappa B Kinase; D016328:NF-kappa B; D017346:Protein Serine-Threonine Kinases; D011993:Recombinant Fusion Proteins; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; D020931:ran GTP-Binding Protein; D000097782:NF-kappaB-Inducing Kinase; C496559:CHUK protein, human; C496560:IKBKB protein, human; C496565:IKBKE protein, human