Reconstitution of the antagonistic effect between C1orf112/FIRRM-FIGNL1 and BRCA2 on RAD51 filament stabilization
Summary: C1orf112/FIRRM is a recently identified DNA damage repair factor that regulates RAD51 in homologous recombination through interacting with the anti-recombinase FIGNL1. Here, we describe steps for purifying C1orf112/FIRRM, FIGNL1, miBRCA2, and RAD51 proteins from Escherichia coli or Saccharo...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2024-03-01
|
| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S266616672300758X |
| _version_ | 1797382889860497408 |
|---|---|
| author | Zenan Zhou Han Yang Xinxin Liang Tao Zhou Qixiang Liu Jiadong Wang Weibin Wang |
| author_facet | Zenan Zhou Han Yang Xinxin Liang Tao Zhou Qixiang Liu Jiadong Wang Weibin Wang |
| author_sort | Zenan Zhou |
| collection | DOAJ |
| description | Summary: C1orf112/FIRRM is a recently identified DNA damage repair factor that regulates RAD51 in homologous recombination through interacting with the anti-recombinase FIGNL1. Here, we describe steps for purifying C1orf112/FIRRM, FIGNL1, miBRCA2, and RAD51 proteins from Escherichia coli or Saccharomyces cerevisiae cells. We then detail procedures for reconstituting the disassembly of RAD51 filament by C1orf112/FIRRM-FIGNL1 in vitro and the antagonistic effect between C1orf112/FIRRM-FIGNL1 and miBRCA2 on RAD51 filament stabilization.For complete details on the use and execution of this protocol, please refer to Zhou et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-08T21:11:56Z |
| format | Article |
| id | doaj.art-f7efd3d19561463fb47a574db242dcd7 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-08T21:11:56Z |
| publishDate | 2024-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-f7efd3d19561463fb47a574db242dcd72023-12-22T05:34:18ZengElsevierSTAR Protocols2666-16672024-03-0151102791Reconstitution of the antagonistic effect between C1orf112/FIRRM-FIGNL1 and BRCA2 on RAD51 filament stabilizationZenan Zhou0Han Yang1Xinxin Liang2Tao Zhou3Qixiang Liu4Jiadong Wang5Weibin Wang6Department of Radiation Medicine, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, ChinaDepartment of Radiation Medicine, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, ChinaDepartment of Radiation Medicine, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, ChinaDepartment of Radiation Medicine, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, ChinaDepartment of Radiation Medicine, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, ChinaDepartment of Radiation Medicine, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, China; Corresponding authorDepartment of Radiation Medicine, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, China; Corresponding authorSummary: C1orf112/FIRRM is a recently identified DNA damage repair factor that regulates RAD51 in homologous recombination through interacting with the anti-recombinase FIGNL1. Here, we describe steps for purifying C1orf112/FIRRM, FIGNL1, miBRCA2, and RAD51 proteins from Escherichia coli or Saccharomyces cerevisiae cells. We then detail procedures for reconstituting the disassembly of RAD51 filament by C1orf112/FIRRM-FIGNL1 in vitro and the antagonistic effect between C1orf112/FIRRM-FIGNL1 and miBRCA2 on RAD51 filament stabilization.For complete details on the use and execution of this protocol, please refer to Zhou et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S266616672300758XCell BiologyMolecular BiologyProtein Biochemistry |
| spellingShingle | Zenan Zhou Han Yang Xinxin Liang Tao Zhou Qixiang Liu Jiadong Wang Weibin Wang Reconstitution of the antagonistic effect between C1orf112/FIRRM-FIGNL1 and BRCA2 on RAD51 filament stabilization STAR Protocols Cell Biology Molecular Biology Protein Biochemistry |
| title | Reconstitution of the antagonistic effect between C1orf112/FIRRM-FIGNL1 and BRCA2 on RAD51 filament stabilization |
| title_full | Reconstitution of the antagonistic effect between C1orf112/FIRRM-FIGNL1 and BRCA2 on RAD51 filament stabilization |
| title_fullStr | Reconstitution of the antagonistic effect between C1orf112/FIRRM-FIGNL1 and BRCA2 on RAD51 filament stabilization |
| title_full_unstemmed | Reconstitution of the antagonistic effect between C1orf112/FIRRM-FIGNL1 and BRCA2 on RAD51 filament stabilization |
| title_short | Reconstitution of the antagonistic effect between C1orf112/FIRRM-FIGNL1 and BRCA2 on RAD51 filament stabilization |
| title_sort | reconstitution of the antagonistic effect between c1orf112 firrm fignl1 and brca2 on rad51 filament stabilization |
| topic | Cell Biology Molecular Biology Protein Biochemistry |
| url | http://www.sciencedirect.com/science/article/pii/S266616672300758X |
| work_keys_str_mv | AT zenanzhou reconstitutionoftheantagonisticeffectbetweenc1orf112firrmfignl1andbrca2onrad51filamentstabilization AT hanyang reconstitutionoftheantagonisticeffectbetweenc1orf112firrmfignl1andbrca2onrad51filamentstabilization AT xinxinliang reconstitutionoftheantagonisticeffectbetweenc1orf112firrmfignl1andbrca2onrad51filamentstabilization AT taozhou reconstitutionoftheantagonisticeffectbetweenc1orf112firrmfignl1andbrca2onrad51filamentstabilization AT qixiangliu reconstitutionoftheantagonisticeffectbetweenc1orf112firrmfignl1andbrca2onrad51filamentstabilization AT jiadongwang reconstitutionoftheantagonisticeffectbetweenc1orf112firrmfignl1andbrca2onrad51filamentstabilization AT weibinwang reconstitutionoftheantagonisticeffectbetweenc1orf112firrmfignl1andbrca2onrad51filamentstabilization |