MED1, a novel binding partner of BRCA1, regulates homologous recombination and R-loop processing
Abstract Homologous recombination (HR) is a major repair pathway of DNA double-strand breaks and is closely related to carcinogenesis. HR deficiency has been established as a therapeutic target. The aim of this study was to elucidate the functions of a novel HR factor, Mediator complex subunit 1 (ME...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-10-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-21495-8 |
| _version_ | 1798029838247788544 |
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| author | Harunori Honjoh Michihiro Tanikawa Osamu Wada-Hiraike Katsutoshi Oda Hirofumi Inaba Asako Kukita Yoshiko Kawata Misako Kusakabe Saki Tsuchimochi Ayumi Taguchi Yuichiro Miyamoto Kenbun Sone Tetsushi Tsuruga Mayuyo Mori-Uchino Yoko Matsumoto Yutaka Osuga |
| author_facet | Harunori Honjoh Michihiro Tanikawa Osamu Wada-Hiraike Katsutoshi Oda Hirofumi Inaba Asako Kukita Yoshiko Kawata Misako Kusakabe Saki Tsuchimochi Ayumi Taguchi Yuichiro Miyamoto Kenbun Sone Tetsushi Tsuruga Mayuyo Mori-Uchino Yoko Matsumoto Yutaka Osuga |
| author_sort | Harunori Honjoh |
| collection | DOAJ |
| description | Abstract Homologous recombination (HR) is a major repair pathway of DNA double-strand breaks and is closely related to carcinogenesis. HR deficiency has been established as a therapeutic target. The aim of this study was to elucidate the functions of a novel HR factor, Mediator complex subunit 1 (MED1), and its association with BRCA1. Formation of the MED1/BRCA1 complex was examined by immunoprecipitation and GST-pull down assays. The transcription cofactor role of BRCA1 was evaluated using luciferase assays. The roles of MED1 on DNA damage response and HR were analyzed by immunofluorescence and HR assays. R-loop accumulation was analyzed using immunofluorescence. R-loop-induced DNA damage was analyzed by comet assays. Immunoprecipitation and GST-pull down assays demonstrated that MED1 is a novel binding partner of BRCA1 and binds to the BRCT domain. Luciferase assays showed that MED1 potentiated the transcription ability of BRCT by two-fold. In MED1-depleted cells, recruitment of HR genes, such as RPA and γH2AX, to DNA damage sites was severely impaired. HR assays showed that MED1 knockdown significantly decreased HR activity. R-loop nuclear accumulation and R-loop-induced comet tails were observed in MED1-depleted cells. We conclude that the transcription factor MED1 contributes to the regulation of the HR pathway and R-loop processing. |
| first_indexed | 2024-04-11T19:30:54Z |
| format | Article |
| id | doaj.art-70d1c957390a4af6ab20a679cbdd5d2b |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-11T19:30:54Z |
| publishDate | 2022-10-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-70d1c957390a4af6ab20a679cbdd5d2b2022-12-22T04:06:59ZengNature PortfolioScientific Reports2045-23222022-10-0112111210.1038/s41598-022-21495-8MED1, a novel binding partner of BRCA1, regulates homologous recombination and R-loop processingHarunori Honjoh0Michihiro Tanikawa1Osamu Wada-Hiraike2Katsutoshi Oda3Hirofumi Inaba4Asako Kukita5Yoshiko Kawata6Misako Kusakabe7Saki Tsuchimochi8Ayumi Taguchi9Yuichiro Miyamoto10Kenbun Sone11Tetsushi Tsuruga12Mayuyo Mori-Uchino13Yoko Matsumoto14Yutaka Osuga15Department of Obstetrics and Gynecology, Graduate School of Medicine, The University of TokyoDepartment of Obstetrics and Gynecology, Graduate School of Medicine, The University of TokyoDepartment of Obstetrics and Gynecology, Graduate School of Medicine, The University of TokyoDivision of Integrative Genomics, Graduate School of Medicine, The University of TokyoDepartment of Obstetrics and Gynecology, Graduate School of Medicine, The University of TokyoDepartment of Obstetrics and Gynecology, Graduate School of Medicine, The University of TokyoDepartment of Obstetrics and Gynecology, Graduate School of Medicine, The University of TokyoDepartment of Obstetrics and Gynecology, Graduate School of Medicine, The University of TokyoDepartment of Obstetrics and Gynecology, Graduate School of Medicine, The University of TokyoDepartment of Obstetrics and Gynecology, Graduate School of Medicine, The University of TokyoDepartment of Obstetrics and Gynecology, Graduate School of Medicine, The University of TokyoDepartment of Obstetrics and Gynecology, Graduate School of Medicine, The University of TokyoDepartment of Obstetrics and Gynecology, Graduate School of Medicine, The University of TokyoDepartment of Obstetrics and Gynecology, Graduate School of Medicine, The University of TokyoDepartment of Obstetrics and Gynecology, Graduate School of Medicine, The University of TokyoDepartment of Obstetrics and Gynecology, Graduate School of Medicine, The University of TokyoAbstract Homologous recombination (HR) is a major repair pathway of DNA double-strand breaks and is closely related to carcinogenesis. HR deficiency has been established as a therapeutic target. The aim of this study was to elucidate the functions of a novel HR factor, Mediator complex subunit 1 (MED1), and its association with BRCA1. Formation of the MED1/BRCA1 complex was examined by immunoprecipitation and GST-pull down assays. The transcription cofactor role of BRCA1 was evaluated using luciferase assays. The roles of MED1 on DNA damage response and HR were analyzed by immunofluorescence and HR assays. R-loop accumulation was analyzed using immunofluorescence. R-loop-induced DNA damage was analyzed by comet assays. Immunoprecipitation and GST-pull down assays demonstrated that MED1 is a novel binding partner of BRCA1 and binds to the BRCT domain. Luciferase assays showed that MED1 potentiated the transcription ability of BRCT by two-fold. In MED1-depleted cells, recruitment of HR genes, such as RPA and γH2AX, to DNA damage sites was severely impaired. HR assays showed that MED1 knockdown significantly decreased HR activity. R-loop nuclear accumulation and R-loop-induced comet tails were observed in MED1-depleted cells. We conclude that the transcription factor MED1 contributes to the regulation of the HR pathway and R-loop processing.https://doi.org/10.1038/s41598-022-21495-8 |
| spellingShingle | Harunori Honjoh Michihiro Tanikawa Osamu Wada-Hiraike Katsutoshi Oda Hirofumi Inaba Asako Kukita Yoshiko Kawata Misako Kusakabe Saki Tsuchimochi Ayumi Taguchi Yuichiro Miyamoto Kenbun Sone Tetsushi Tsuruga Mayuyo Mori-Uchino Yoko Matsumoto Yutaka Osuga MED1, a novel binding partner of BRCA1, regulates homologous recombination and R-loop processing Scientific Reports |
| title | MED1, a novel binding partner of BRCA1, regulates homologous recombination and R-loop processing |
| title_full | MED1, a novel binding partner of BRCA1, regulates homologous recombination and R-loop processing |
| title_fullStr | MED1, a novel binding partner of BRCA1, regulates homologous recombination and R-loop processing |
| title_full_unstemmed | MED1, a novel binding partner of BRCA1, regulates homologous recombination and R-loop processing |
| title_short | MED1, a novel binding partner of BRCA1, regulates homologous recombination and R-loop processing |
| title_sort | med1 a novel binding partner of brca1 regulates homologous recombination and r loop processing |
| url | https://doi.org/10.1038/s41598-022-21495-8 |
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