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

Autor(es)

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

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo Científico
• Fonte: MUTAGENESIS; v. 36, n. 3, p. 223-236, MAY 2021.
• Citações Web of Science: 0

Resumo

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)

• Processo FAPESP: 16/02040-8 - Analise funcional de mutações em nek1 e nek7 em câncer e identificação dos seus substratos fisiológicos
• Beneficiário: Mariana Bonjiorno Martins
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 14/15982-6 - Consequências de deficiências de reparo de lesões no genoma
• Beneficiário: Carlos Frederico Martins Menck
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 17/21067-7 - Análise do papel de NEK1 na regulação da resposta à danos de DNA nuclear e mitocondrial
• Beneficiário: Mariana Bonjiorno Martins
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Pós-Doutorado
• Processo FAPESP: 17/03489-1 - De estudos funcionais à busca de novos inibidores anticâncer: explorando cinases reguladores do ciclo celular da família de NEK humana
• Beneficiário: Jörg Kobarg
• Modalidade de apoio: Auxílio à Pesquisa - Temático