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.

中文翻译：

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NEK1缺乏影响线粒体功能和关键DNA修复途径的转录组

以前的研究表明 NIMA 相关激酶 1 (NEK1) 在调节 DNA 损伤检查点和 DNA 修复能力中的重要作用。为了广泛评估 NEK1 对基因毒性应激和线粒体功能的贡献，我们表征了 NEK1 CRISPR 敲除 (KO) 和野生型 (WT) HAP1 细胞的几种相关表型。我们的研究表明，NEK1 KO 细胞导致细胞凋亡增加和对烷化剂甲磺酸甲酯、放射模拟博莱霉素和 UVC 光的超敏性增加，但对交联剂顺铂的抗性增加。NEK1 KO 细胞中的线粒体功能也发生了改变，表现为线粒体自噬减少、总线粒体增加、活性氧水平升高、复合物 I 活性受损和线粒体 DNA 损伤增加。RNA-seq 转录组分析结合定量实时 PCR 研究比较了 NEK1 KO 细胞和 NEK1 过表达细胞，揭示了参与 DNA 修复途径的基因的表达，例如碱基切除修复、核苷酸切除修复和双链断裂修复。以可能影响基因毒素抗性的方式改变。总之，我们的研究强调并进一步支持 NEK1 作为一个中枢信号激酶响应 DNA 损伤、调节 DNA 修复能力、线粒体活性和细胞命运决定。以可能影响基因毒素抗性的方式改变。总之，我们的研究强调并进一步支持 NEK1 作为一个中枢信号激酶响应 DNA 损伤、调节 DNA 修复能力、线粒体活性和细胞命运决定。以可能影响基因毒素抗性的方式改变。总之，我们的研究强调并进一步支持 NEK1 作为一个中枢信号激酶响应 DNA 损伤、调节 DNA 修复能力、线粒体活性和细胞命运决定。