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Systematic investigation of BRCA1-A, -B, and -C complexes and their functions in DNA damage response and DNA repair

Oncogene volume 43, pages 2621–2634 (2024)Cite this article

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Abstract

BRCA1, a breast cancer susceptibility gene, has emerged as a central mediator that brings together multiple signaling complexes in response to DNA damage. The A, B, and C complexes of BRCA1, which are formed based on their phosphorylation-dependent interactions with the BRCA1-C-terminal domains, contribute to the roles of BRCA1 in DNA repair and cell cycle checkpoint control. However, their functions in DNA damage response remain to be fully appreciated. Specifically, there has been no systematic investigation of the roles of BRCA1-A, -B, and -C complexes in the regulation of BRCA1 localization and functions, in part because of cellular lethality associated with loss of CtIP protein, which is an essential component in BRCA1-C complex. To systematically investigate the functions of these complexes in DNA damage response, we depleted a key component in each of these complexes. We used the degradation tag system to inducibly deplete endogenous CtIP and obtained a series of RAP80/FANCJ/CtIP single-, double-, and triple-knockout cells. We showed that loss of BRCA1-B/FANCJ and BRCA1-C/CtIP, but not BRCA1-A/RAP80, resulted in reduced cell proliferation and increased sensitivity to DNA damage. BRCA1-C/CtIP and BRCA1-A/RAP80 were involved in BRCA1 recruitment to sites of DNA damage. However, BRCA1-A/RAP80 was not essential for damage-induced BRCA1 localization. Instead, RAP80/H2AX and CtIP have redundant roles in BRCA1 recruitment. Altogether, our systematic analysis uncovers functional differences between BRCA1-A, -B, and -C complexes and provides new insights into the roles of these BRCA1-associated protein complexes in DNA damage response and DNA repair.

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Fig. 1: FANCJ depletion reduces cell viability and enhances cellular sensitivity to DNA damage-inducing agents.
Fig. 2: CtIP is essential for cell viability.
Fig. 3: CtIP, RAP80, and FANCJ regulate BRCA1 functions in response to DNA damage.
Fig. 4: CtIP and H2AX represent two independent pathways involved in BRCA1 recruitment.

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Acknowledgements

We thank all members of Dr. Chen’s laboratory for their help and constructive discussions and the North Campus Flow Cytometry and Cellular Imaging Core Facility, DNA Analysis Core Facility at the University of Texas MD Anderson Cancer (CA016672) for their technical support. We thank Sarah Bronson and Ann Sutton at the Research Medical Library for Editing Services. We also thank Drs. Richard Baer and Sharon Cantor respectively for providing CtIP and FANCJ antibodies.

Funding

This work was supported in part by internal funds that were available to JC. JC also received support from the Pamela and Wayne Garrison Distinguished Chair in Cancer Research, Cancer Prevention & Research Institute of Texas (awards RP160667 and RP180813 to JC), and NIH (CA210929, CA216911, and CA216437 to JC; P01 CA193124, project 4, to JC; and more recently, CA274234 and CA275712 to JC).

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  1. These authors contributed equally: Siting Li, Mengfan Tang.

Authors and Affiliations

  1. Department of Experimental Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    Siting Li, Mengfan Tang, Yun Xiong, Xu Feng, Chao Wang, Litong Nie, Min Huang, Huimin Zhang, Ling Yin, Dandan Zhu, Chang Yang, Tiantian Ma & Junjie Chen

  2. Department of Immunology, School of Medicine, Nanjing University of Chinese Medicine, Nanjing, China

    Mengfan Tang

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Contributions

Siting Li, Mengfan Tang, and Junjie Chen conceived the project. Siting Li, Mengfan Tang, Litong Nie, Min Huang, Huimin Zhang, Ling Yin, Dandan Zhu, Chang Yang, and Tiantian Ma performed the experiments. Yun Xiong, Xu Feng, and Chao Wang helped with the mass spectrometry experiments and CRISPR screen data analysis, and Siting Li and Junjie Chen wrote the manuscript with input from all authors.

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Correspondence to Junjie Chen.

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Li, S., Tang, M., Xiong, Y. et al. Systematic investigation of BRCA1-A, -B, and -C complexes and their functions in DNA damage response and DNA repair. Oncogene 43, 2621–2634 (2024). https://doi.org/10.1038/s41388-024-03108-y

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