NEK1 deficiency affects mitochondrial functions and the transcriptome of key DNA repair pathways

Author(s)

Martins, Mariana Bonjiorno ^[1] ; Perez, Arina Marina ^[2] ; Bohr, Vilhelm A. ^[2] ; Wilson, III, David M. ; Kobarg, Jorg ^{[1, 3]}

Affiliation

^[1] Univ Estadual Campinas, Inst Biol, Dept Bioquim & Biol Tecidual, Campinas, SP - Brazil

^[2] NIA, Lab Mol Gerontol, NIH, Baltimore, MD 21224 - USA

^[3] Univ Estadual Campinas, Caculdade Ciencias Farmaceut, Campinas, SP - Brazil

• Document type: Journal article
• Source: MUTAGENESIS; v. 36, n. 3, p. 223-236, MAY 2021.
• Web of Science Citations: 0

Abstract

Previous studies have indicated important roles for NIMA-related kinase 1 (NEK1) in modulating DNA damage checkpoints and DNA repair capacity. To broadly assess the contributions of NEK1 to genotoxic stress and mitochondrial functions, we characterised several relevant phenotypes of NEK1 CRISPR knockout (KO) and wild-type (WT) HAP1 cells. Our studies revealed that NEK1 KO cells resulted in increased apoptosis and hypersensitivity to the alkylator methyl methanesulfonate, the radiomimetic bleomycin and UVC light, yet increased resistance to the crosslinker cisplatin. Mitochondrial functionalities were also altered in NEK1 KO cells, with phenotypes of reduced mitophagy, increased total mitochondria, elevated levels of reactive oxygen species, impaired complex I activity and higher amounts of mitochondrial DNA damage. RNA-seq transcriptome analysis coupled with quantitative real-time PCR studies comparing NEK1 KO cells with NEK1 overexpressing cells revealed that the expression of genes involved in DNA repair pathways, such as base excision repair, nucleotide excision repair and double-strand break repair, are altered in a way that might influence genotoxin resistance. Together, our studies underline and further support that NEK1 serves as a hub signalling kinase in response to DNA damage, modulating DNA repair capacity, mitochondrial activity and cell fate determination. (AU)

• FAPESP's process: 16/02040-8 - FUNCTIONAL ANALYSIS OF MUTATIONS IN NEK1 AND NEK7 IN CANCER AND IDENTIFICATION OF THEIR PHYSIOLOGICAL SUBSTRATES
• Grantee: Mariana Bonjiorno Martins
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 14/15982-6 - Consequences of repair deficiencies in damaged genome
• Grantee: Carlos Frederico Martins Menck
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 17/21067-7 - Analysis of the role of NEK1 in the regulation of the response to nuclear and mitochondrial DNA damage
• Grantee: Mariana Bonjiorno Martins
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• FAPESP's process: 17/03489-1 - From functional studies to searching for new inhibitors for cancer: exploring kinases that regulate the cell cycle of the human NEK family
• Grantee: Jörg Kobarg
• Support Opportunities: Research Projects - Thematic Grants