Reconstitution of the antagonistic effect between C1orf112/FIRRM-FIGNL1 and BRCA2 on RAD51 filament stabilization

Zenan Zhou; Han Yang; Xinxin Liang; Tao Zhou; Qixiang Liu; Jiadong Wang; Weibin Wang

doi:10.1016/j.xpro.2023.102791

Reconstitution of the antagonistic effect between C1orf112/FIRRM-FIGNL1 and BRCA2 on RAD51 filament stabilization

STAR Protoc. 2024 Mar 15;5(1):102791. doi: 10.1016/j.xpro.2023.102791. Epub 2023 Dec 20.

Authors

Zenan Zhou¹, Han Yang¹, Xinxin Liang¹, Tao Zhou¹, Qixiang Liu¹, Jiadong Wang², Weibin Wang³

Affiliations

¹ Department of Radiation Medicine, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, China.
² Department of Radiation Medicine, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, China. Electronic address: wangjd@bjmu.edu.cn.
³ Department of Radiation Medicine, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, China. Electronic address: weibinwang@bjmu.edu.cn.

Abstract

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.

Keywords: Cell Biology; Molecular Biology; Protein Biochemistry.

MeSH terms

DNA Repair
Homologous Recombination
Proteins* / genetics
Rad51 Recombinase* / genetics

Substances

Proteins
Rad51 Recombinase