Elp1 facilitates RAD51-mediated homologous recombination repair via translational regulation

Abstract 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 impl...

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Autores principales:	Wei-Ting Chen, Huan-Yi Tseng, Chung-Lin Jiang, Chih-Ying Lee, Peter Chi, Liuh-Yow Chen, Kai-Yin Lo, I-Ching Wang, Fu-Jung Lin
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	Elp1 RAD51 Homologous recombination DNA damage Translational regulation Medicine R
Acceso en línea:	https://doaj.org/article/a3c039a92bb749de84509f151dd98baf
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spelling	oai:doaj.org-article:a3c039a92bb749de84509f151dd98baf2021-11-28T12:14:17ZElp1 facilitates RAD51-mediated homologous recombination repair via translational regulation10.1186/s12929-021-00773-z1423-0127https://doaj.org/article/a3c039a92bb749de84509f151dd98baf2021-11-01T00:00:00Zhttps://doi.org/10.1186/s12929-021-00773-zhttps://doaj.org/toc/1423-0127Abstract 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.Wei-Ting ChenHuan-Yi TsengChung-Lin JiangChih-Ying LeePeter ChiLiuh-Yow ChenKai-Yin LoI-Ching WangFu-Jung LinBMCarticleElp1RAD51Homologous recombinationDNA damageTranslational regulationMedicineRENJournal of Biomedical Science, Vol 28, Iss 1, Pp 1-19 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Elp1 RAD51 Homologous recombination DNA damage Translational regulation Medicine R
spellingShingle	Elp1 RAD51 Homologous recombination DNA damage Translational regulation Medicine R Wei-Ting Chen Huan-Yi Tseng Chung-Lin Jiang Chih-Ying Lee Peter Chi Liuh-Yow Chen Kai-Yin Lo I-Ching Wang Fu-Jung Lin Elp1 facilitates RAD51-mediated homologous recombination repair via translational regulation
description	Abstract 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.
format	article
author	Wei-Ting Chen Huan-Yi Tseng Chung-Lin Jiang Chih-Ying Lee Peter Chi Liuh-Yow Chen Kai-Yin Lo I-Ching Wang Fu-Jung Lin
author_facet	Wei-Ting Chen Huan-Yi Tseng Chung-Lin Jiang Chih-Ying Lee Peter Chi Liuh-Yow Chen Kai-Yin Lo I-Ching Wang Fu-Jung Lin
author_sort	Wei-Ting Chen
title	Elp1 facilitates RAD51-mediated homologous recombination repair via translational regulation
title_short	Elp1 facilitates RAD51-mediated homologous recombination repair via translational regulation
title_full	Elp1 facilitates RAD51-mediated homologous recombination repair via translational regulation
title_fullStr	Elp1 facilitates RAD51-mediated homologous recombination repair via translational regulation
title_full_unstemmed	Elp1 facilitates RAD51-mediated homologous recombination repair via translational regulation
title_sort	elp1 facilitates rad51-mediated homologous recombination repair via translational regulation
publisher	BMC
publishDate	2021
url	https://doaj.org/article/a3c039a92bb749de84509f151dd98baf
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_version_	1718408158608621568

Elp1 facilitates RAD51-mediated homologous recombination repair via translational regulation

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