Characterization of NEK10 proteomic interactions under genotoxic stress

ABSTRACT: Genetic studies in Aspergillus nidulans identified a family of protein kinases (NIMA- Never in mitosis gene A) crucial for cell cycle progression in fungus. Highly conserved in mammals, various members of NEKs (Nima-related kinases) cross-function in signaling networks depicted in at least one of the following main biological processes: mitotic events, DNA damage repair and response and in cilliagenesis. Recent studies have shown new functions for NEK1, 2, 4 and 5 in mitochondria respiration, apoptosis and protein complexes. NEK10 is the least studied member of the Neks, nevertheless NEK10 has been reported in Neks recurrent functions: G2/M control, centrosomes and cilliagenesis. Besides, the downregulation of Nek10 was associated with poor prognosis and aggressive breast tumors. Given that Neks function relies on protein signaling networks, we investigated Nek10 protein interacting partners and Nek10´s biological roles through IP-MS/MS. Our mass spectrometry data identified new interactions for Nek10 proteomics including mitochondria functional genes: SIRT3, ATAD3A, ATAD3B and OAT. The number of mitochondrial partners was augmented more than 3-fold after zeocin treatment: FKBP4, TXN, PFDN2, ATAD3B, MRPL12, ATP5J, DUT, YWHAE, CS, SIRT3, HSPA9, PDHB, GLUD1, DDX3X, and APEX1. We focused on exerting Nek10 function on the mitochondria and we first confirmed Nek10 localization in the mitochondria and its interaction with mitochondria proteins. To extend our understanding in Nek10 roles, we depleted Nek10 in mammalian cell lines further denoting mitochondria-related phenotypes such as inhibition of mitochondrial respiration, loss of mtDNA amplification and defects on mitochondria morphology. Our proteomics and functional data contributes to the first evidences of Nek10 role in mitochondria and our overall work drive Nek10 unknown roads towards the avenues of modulating mechanisms of diseases.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell

SUBMITTER: PRISCILA SLEPICKA  

LAB HEAD: Jörg Kobarg

PROVIDER: PXD018294 | Pride | 2020-05-06

REPOSITORIES: Pride