title: Characterization of the human NEK7 interactome suggests catalytic and regulatory properties distinct from those of NEK6.
abstract: Human NEK7 is a regulator of cell division and plays an important role in growth and survival of mammalian cells. Human NEK6 and NEK7 are closely related, consisting of a conserved C-terminal catalytic domain and a nonconserved and disordered N-terminal regulatory domain, crucial to mediate the interactions with their respective proteins. Here, in order to better understand NEK7 cellular functions, we characterize the NEK7 interactome by two screening approaches: one using a yeast two-hybrid system and the other based on immunoprecipitation followed by mass spectrometry analysis. These approaches led to the identification of 61 NEK7 interactors that contribute to a variety of biological processes, including cell division. Combining additional interaction and phosphorylation assays from yeast two-hybrid screens, we validated CC2D1A, TUBB2B, MNAT1, and NEK9 proteins as potential NEK7 interactors and substrates. Notably, endogenous RGS2, TUBB, MNAT1, NEK9, and PLEKHA8 localized with NEK7 at key sites throughout the cell cycle, especially during mitosis and cytokinesis. Furthermore, we obtained evidence that the closely related kinases NEK6 and NEK7 do not share common interactors, with the exception of NEK9, and display different modes of protein interaction, depending on their N- and C-terminal regions, in distinct fashions. In summary, our work shows for the first time a comprehensive NEK7 interactome that, combined with functional in vitro and in vivo assays, suggests that NEK7 is a multifunctional kinase acting in different cellular processes in concert with cell division signaling and independently of NEK6.
authors.affiliation: Laboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e Materiais , Campinas, São Paulo, Brazil.
authors.forename: Edmarcia Elisa
authors.initials: EE
authors.lastname: de Souza
authors.forename: Gabriela Vaz
authors.initials: GV
authors.lastname: Meirelles
authors.forename: Bárbara Biatriz
authors.initials: BB
authors.lastname: Godoy
authors.forename: Arina Marina
authors.initials: AM
authors.lastname: Perez
authors.forename: Juliana Helena Costa
authors.initials: JH
authors.lastname: Smetana
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Doxsey
authors.forename: Mark E
authors.initials: ME
authors.lastname: McComb
authors.forename: Catherine E
authors.initials: CE
authors.lastname: Costello
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Whelan
authors.forename: Jörg
authors.initials: J
authors.lastname: Kobarg
mesh_terms: D002453:Cell Cycle / Q000502:physiology; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D000072379:NIMA-Related Kinases; D011485:Protein Binding; D060066:Protein Interaction Maps / Q000502:physiology*; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000378:metabolism*; D040901:Proteomics; D020798:Two-Hybrid System Techniques
chemical_list: C459456:NEK6 protein, human; C512685:NEK7 protein, human; D000072379:NIMA-Related Kinases; D017346:Protein Serine-Threonine Kinases
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