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K63-linked ubiquitination of FANCG is required for its association with the Rap80-BRCA1 complex to modulate homologous recombination repair of DNA interstand crosslinks

Oncogene volume 34, pages 2867–2878 (2015)Cite this article

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Abstract

DNA interstrand crosslinks (ICLs) are extremely deleterious lesions that are repaired by homologous recombination (HR) through coordination of Fanconi anemia (FA) proteins and breast cancer susceptibility gene 1 (BRCA1) product, but the exact role these proteins have remains unclear. Here we report that FANCG was modified by the addition of lysine63-linked polyubiquitin chains (K63Ub) in response to DNA damage. We show that FANCG K63Ub was dispensable for monoubiquitination of FANCD2, but was required for FANCG to interact with the Rap80-BRCA1 (receptor-associated protein 80-BRCA1) complex for subsequent modulation of HR repair of ICLs induced by mitomycin C. Mutation of three lysine residues within FANCG to arginine (K182, K258 and K347, 3KR) reduced FANCG K63Ub modification, as well as its interaction with the Rap80-BRCA1 complex, and therefore impeded HR repair. In addition, we demonstrated that K63Ub-modified FANCG was deubiquitinated by BRCC36 complex in vitro and in vivo. Inhibition of BRCC36 resulted in increased K63Ub modification of FANCG. Taken together, our results identify a new role of FANCG in HR repair of ICL through K63Ub-mediated interaction with the Rap80-BRCA1 complex.

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Acknowledgements

This work was supported by Beijing Natural Science Foundation (Grant 7122099); Natural Science Foundation of China (Grant 81272915); the Research Fund for the Doctoral Program of Higher Education (20110001110013); the Project Sponsored by the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry (JWSL44-8); 985 Program, Ministry of Education of China.

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Authors and Affiliations

  1. Department of Cell Biology, School of Basic Medical Sciences, Peking University, Beijing, China

    B Zhu, K Yan, L Li, M Lin, S Zhang, H Yang & G Shao

  2. Institute of Systems Biology, Peking University, Beijing, China

    B Zhu & G Shao

  3. Center of Medical and Health Analysis, Peking University, Beijing, China

    Q He

  4. School of Medicine, Shenzhen University, Shenzhen, Guangdong, China

    D Zheng

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  1. B Zhu
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Zhu, B., Yan, K., Li, L. et al. K63-linked ubiquitination of FANCG is required for its association with the Rap80-BRCA1 complex to modulate homologous recombination repair of DNA interstand crosslinks. Oncogene 34, 2867–2878 (2015). https://doi.org/10.1038/onc.2014.229

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