@article{ab3583f49e1d445aaef37bfbd7eb382a,
title = "Nuclease-independent functions of RAG1 direct distinct DNA damage responses in B cells",
abstract = "Developing B cells generate DNA double-stranded breaks (DSBs) to assemble immunoglobulin receptor (Ig) genes necessary for the expression of a mature B cell receptor. These physiologic DSBs are made by the RAG endonuclease, which is comprised of the RAG1 and RAG2 proteins. In pre-B cells, RAG-mediated DSBs activate the ATM kinase to coordinate canonical and non-canonical DNA damage responses (DDR) that trigger DSB repair and B cell developmental signals, respectively. Whether this broad cellular response is distinctive to RAG DSBs is poorly understood. To delineate the factors that direct DDR signaling in B cells, we express a tetracycline-inducible Cas9 nuclease in Rag1-deficient pre-B cells. Both RAG- and Cas9-mediated DSBs at Ig genes activate canonical DDR. In contrast, RAG DSBs, but not Cas9 DSBs, induce the non-canonical DDR-dependent developmental program. This unique response to RAG DSBs is, in part, regulated by non-core regions of RAG1. Thus, B cells trigger distinct cellular responses to RAG DSBs through unique properties of the RAG endonuclease that promotes activation of B cell developmental programs.",
keywords = "B cell development, Cas9, DNA breaks, DNA damage response, RAG",
author = "Rachel Johnston and Brendan Mathias and Crowley, {Stephanie J.} and Schmidt, {Haley A.} and White, {Lynn S.} and Nima Mosammaparast and Green, {Abby M.} and Bednarski, {Jeffrey J.}",
note = "Funding Information: This work was supported by the National Institutes of Health grants K08AI102946 (JJB), R56AI153234 (JJB), R01AI173077 (JJB), R01CA227001 (NM), R01CA193318 (NM), P01CA092584 (NM), and 5T32AI007163 (SJC). JJB was supported by the Foundation for Barnes‐Jewish Hospital Cancer Frontier Fund, Barnard Trust, American Society of Hematology, Gabrielle's Angel Foundation, St. Louis Children's Hospital Foundation, and the Children's Discovery Institute. RJ was supported by a training grant through the Alvin J. Siteman Cancer Center. AMG was supported by the Department of Defense (CA200867) and the Children's Discovery Institute. NM was supported by the Centene Personalized Medicine Initiative, the American Cancer Society (RSG‐18‐156‐01‐DMC), the Barnard Foundation, and the Alvin J. Siteman Cancer Center Investment Program. The visual synopsis was created with BioRender.com . Funding Information: This work was supported by the National Institutes of Health grants K08AI102946 (JJB), R56AI153234 (JJB), R01AI173077 (JJB), R01CA227001 (NM), R01CA193318 (NM), P01CA092584 (NM), and 5T32AI007163 (SJC). JJB was supported by the Foundation for Barnes-Jewish Hospital Cancer Frontier Fund, Barnard Trust, American Society of Hematology, Gabrielle's Angel Foundation, St. Louis Children's Hospital Foundation, and the Children's Discovery Institute. RJ was supported by a training grant through the Alvin J. Siteman Cancer Center. AMG was supported by the Department of Defense (CA200867) and the Children's Discovery Institute. NM was supported by the Centene Personalized Medicine Initiative, the American Cancer Society (RSG-18-156-01-DMC), the Barnard Foundation, and the Alvin J. Siteman Cancer Center Investment Program. The visual synopsis was created with BioRender.com. Publisher Copyright: {\textcopyright} 2022 The Authors.",
year = "2023",
month = jan,
day = "9",
doi = "10.15252/embr.202255429",
language = "English",
volume = "24",
journal = "EMBO Reports",
issn = "1469-221X",
number = "1",
}