https://scholars.lib.ntu.edu.tw/handle/123456789/589896
標題: | Elp1 facilitates RAD51-mediated homologous recombination repair via translational regulation | 作者: | WEI-TING CHEN Tseng, Huan Yi Jiang, Chung Lin Lee, Chih Ying Chi, Peter Chen, Liuh Yow KAI-YIN LO Wang, I. Ching |
關鍵字: | DNA damage | Elp1 | Homologous recombination | RAD51 | Translational regulation;DNA damage; Elp1; Homologous recombination; RAD51; Translational regulation | 公開日期: | 1-十二月-2021 | 出版社: | BioMed Central Ltd | 卷: | 28 | 期: | 1 | 起(迄)頁: | Article number 81 | 來源出版物: | Journal of Biomedical Science | 摘要: | Background: RAD51-dependent homologous recombination (HR) is one of the most important pathways for repairing DNA double-strand breaks (DSBs), and its regulation is crucial to maintain genome integrity. Elp1 gene encodes IKAP/ELP1, a core subunit of the Elongator complex, which has been implicated in translational regulation. However, how ELP1 contributes to genome maintenance is unclear. Methods: To investigate the function of Elp1, Elp1-deficient mouse embryonic fibroblasts (MEFs) were generated. Metaphase chromosome spreading, immunofluorescence, and comet assays were used to access chromosome abnormalities and DSB formation. Functional roles of Elp1 in MEFs were evaluated by cell viability, colony forming capacity, and apoptosis assays. HR-dependent DNA repair was assessed by reporter assay, immunofluorescence, and western blot. Polysome profiling was used to evaluate translational efficiency. Differentially expressed proteins and signaling pathways were identified using a label-free liquid chromatography–tandem mass spectrometry (LC–MS/MS) proteomics approach. Results: Here, we report that Elp1 depletion enhanced genomic instability, manifested as chromosome breakage and genotoxic stress-induced genomic DNA fragmentation upon ionizing radiation (IR) exposure. Elp1-deficient cells were hypersensitive to DNA damage and exhibited impaired cell proliferation and defective HR repair. Moreover, Elp1 depletion reduced the formation of IR-induced RAD51 foci and decreased RAD51 protein levels. Polysome profiling analysis revealed that ELP1 regulated RAD51 expression by promoting its translation in response to DNA damage. Notably, the requirement for ELP1 in DSB repair could be partially rescued in Elp1-deficient cells by reintroducing RAD51, suggesting that Elp1-mediated HR-directed repair of DSBs is RAD51-dependent. Finally, using proteome analyses, we identified several proteins involved in cancer pathways and DNA damage responses as being differentially expressed upon Elp1 depletion. Conclusions: Our study uncovered a molecular mechanism underlying Elp1-mediated regulation of HR activity and provides a novel link between translational regulation and genome stability. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/589896 | ISSN: | 10217770 | DOI: | 10.1186/s12929-021-00773-z | SDG/關鍵字: | protein Rad1; Ikbkap protein, mouse; Rad51 protein; Rad51 protein, mouse; signal peptide; animal cell; animal tissue; apoptosis; Article; cell viability; chromosome aberration; chromosome breakage; colony formation; comet assay; controlled study; DNA damage; DNA repair; Elp1 gene; embryo; fibroblast; gene; gene function; gene silencing; genomic instability; genotoxicity; immunofluorescence; liquid chromatography-mass spectrometry; metaphase chromosome; mouse; nonhuman; proteomics; real time polymerase chain reaction; recombination repair; signal transduction; staining; translation regulation; Western blotting; animal; chromosome breakage; genetics; metabolism; protein synthesis; recombination repair; Animals; Chromosome Breakage; DNA Damage; Fibroblasts; Genomic Instability; Intracellular Signaling Peptides and Proteins; Mice; Protein Biosynthesis; Rad51 Recombinase; Recombinational DNA Repair |
顯示於: | 生化科技學系 |
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