Human NIMA-related kinase 6 is one of the Fe65 WW domain binding proteins.
The Aspergillus nidulans protein NIMA (never in mitosis, gene A) is a protein kinase required for initiation of mitosis, whereas its inactivation is necessary for mitotic exit. Here, we present evidence that human Nek6 is associated with Fe65. Based on the presence of Fe65 WW domain binding motifs ((267)PPLP(270)) in the Nek6 catalytic domain, we observed that Nek6 interacts physically with Fe65 both in vivo and in vitro, using a pull-down approach. Additionally, we detected co-localization of Nek6 and Fe65 via confocal microscopy. Co-localization of Nek6 and Fe65 was disrupted by mutation of the WW domain binding motifs ((267)PPLP(270)). Finally, when transient transfection assays were performed, interaction of Nek6 (wt) with Fe65 induced substantial cell apoptosis, whereas interaction using the Nek6 pplp mutant ((267)PPLP(270) changes (267)APVA(270)) did not. Thus, our observations indicated that Nek6 binds to Fe65 through its (267)PPLP(270) motif and that the protein-protein interaction between Nek6 and Fe65 regulates their subcellular localization and cell apoptosis.
Lee
Eun Jeoung
EJ
School of Science Education, Chungbuk National University, Gaeshin-dong, Heungdok-gu, Chongju, Chungbuk 361-763, Republic of Korea.
Hyun
Sung Hee
SH
Chun
Jaesun
J
Kang
Sang Sun
SS
D000818:Animals; D017209:Apoptosis; D001233:Aspergillus nidulans / Q000378:metabolism*; D019556:COS Cells; D002453:Cell Cycle; D002522:Chlorocebus aethiops; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D008938:Mitosis; D016297:Mutagenesis, Site-Directed; D000072379:NIMA-Related Kinases; D009419:Nerve Tissue Proteins / Q000502:physiology*; D009687:Nuclear Proteins / Q000502:physiology*; D025941:Protein Interaction Mapping; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000502:physiology*; D017434:Protein Structure, Tertiary; D015398:Signal Transduction
C091042:APBB1 protein, human; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D049452:Green Fluorescent Proteins; D000072379:NIMA-Related Kinases; C442331:NIMA-related kinase 6; D017346:Protein Serine-Threonine Kinases