Analysis of the involvement of Nek7 protein in DDR through proteomic and phosphoproteomic studies using SILAC assay

Abstract

Nek7 protein belongs to the NIMA-related kinases or "Neks" family, that comprises a family of eleven proteins named Nek1 to Nek 11 which have been functionally associated to mitosis, cilium regulation and DNA damage response (DDR) (Meirelles et al, 2014). Recent studies showed the involvement of Nek7 in centrosome duplication mechanism. Moreover, is known that Nek7 is over-expressed in breast, colorectal, laryngeal, and lung cancer as well as in non-Hodgin lymphoma. In addition, although we have been connected Nek7 to DDR functions using in silico analysis (Meirelles et al, 2014), the molecular mechanisms underlying the role of Nek7 in DDR are not known. Our recent studies show, for the first time, evidences of the involvement of Nek7 in DDR when cells are irradiated with UV light (in preparation Perez, AM et al).Thus, the employment of functional studies is necessary to clarify the function of Nek7 in carcinogenesis, and its role in the signaling pathway of cell cycle and DDR. Thus, we intend to unveil the signaling pathways related to DNA damage response mechanisms in which Nek7 may be involved. To achieve this objective we will perform proteomics and phosphoproteomic assays using Nek7 wild-type and their kinase dead mutants expressed in normal cells and cells under treatment with different genotoxic agents. This approach will help us to elucidate the molecular mechanisms underlying the role of Nek7 in the regulation of cell cycle and DNA damage response. (AU)