Protein kinase PKN1 associates with TRAF2 and is involved in TRAF2-NF-kappaB signaling pathway.
PKN1 is a fatty acid and Rho-activated serine/threonine protein kinase whose catalytic domain is highly homologous to protein kinase C (PKC) family. In yeast two-hybrid screening for PKN1 binding proteins, we identified tumor necrosis factor alpha (TNFalpha) receptor-associated factor 2 (TRAF2). TRAF2 is one of the major mediators of TNF receptor superfamily transducing TNF signal to various functional targets, including activation of NF-kappaB, JNK, and apoptosis. FLAG-tagged PKN1 was co-immunoprecipitated with endogenous TRAF2 from HEK293 cell lysate, and in vitro binding assay using the deletion mutants of TRAF2 showed that PKN1 directly binds to the TRAF domain of TRAF2. PKN1 has the TRAF2-binding consensus sequences PXQX (S/T) at amino acid residues 580-584 (PIQES), and P580AQ582A mutant was not co-immunoprecipitated with TRAF2. Furthermore, the reduced expression of PKN1 by RNA interference (RNAi) down-regulated TRAF2-induced NF-kappaB activation in HEK293T cells. These results suggest that PKN1 is involved in TRAF2-NF-kappaB signaling pathway.
Gotoh
Yusuke
Y
Graduate School of Science and Technology, Kobe University, 657-8501, Kobe, Japan.
Oishi
Kumiko
K
Shibata
Hideki
H
Yamagiwa
Akiko
A
Isagawa
Takayuki
T
Nishimura
Tamako
T
Goyama
Emiko
E
Takahashi
Mikiko
M
Mukai
Hideyuki
H
Ono
Yoshitaka
Y
D000595:Amino Acid Sequence; D002460:Cell Line; D015536:Down-Regulation; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism / Q000502:physiology*; D011233:Precipitin Tests; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D011506:Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D034622:RNA Interference; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D047992:TNF Receptor-Associated Factor 2; D014162:Transfection; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics
D016328:NF-kappa B; D011506:Proteins; D011994:Recombinant Proteins; D047992:TNF Receptor-Associated Factor 2; D008156:Luciferases; D017346:Protein Serine-Threonine Kinases