Nek2 is a mammalian protein kinase structurally homologous to Aspergillus NIMA. We previously observed that the Nek2 protein was localized in multiple sites within a cell in a cell cycle stage-specific manner. Such dynamic behavior of Nek2 allowed us to propose that Nek2 may be a mitotic regulator that is involved in diverse cell cycle events. To better understand the cellular processes in which Nek2 participates, we carried out yeast two-hybrid screening and isolated Nek2-Interacting Protein 1 (NIP1), which has been also named as XB51 and NECAB3. Physical interactions of Nek2 with NIP1 were confirmed. In fact, Nek2 can phosphorylate NIP1 in vivo. Immunostaining experiments revealed that NIP1 is a Golgi protein. These results propose a possible involvement of Nek2 in biological processes of the Golgi body, perhaps in relation to the inheritance of Golgi during mitosis or to cell cycle stage-specific regulation of exocytosis.