The DNA Damage Response Regulates RAG1/2 Expression in Pre–B Cells through ATM-FOXO1 Signaling

Ochodnicka‐Mackovicova, Katarina; Bahjat, Mahnoush; Maas, Chiel; Veen, Amélie van der; Bloedjes, Timon A.; Bruin, Alexander M. de; Andel, Harmen van; Schrader, Carol E.; Hendriks, Rudi W.; Verhoeyen, Els; Bende, Richard J.; Noesel, Carel J. M. van; Guikema, Jeroen E. J.

doi:10.4049/jimmunol.1501989

Cited by 30 publications

(32 citation statements)

References 62 publications

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“…Previous studies reported that the Rag1 and Rag2 proteins each have a half-life of ~10–12 minutes (29, 47, 48). Considering these reports and our observed kinetics and extents of Rag1 and Rag2 mRNA loss in response to IR, one would predict appreciable decreased levels of Rag1 and Rag2 protein at 1 hour after irradiation.…”

Section: Resultsmentioning

confidence: 99%

“…While we were preparing this manuscript, the Guikema lab reported that the DDR response regulates Rag1/Rag2 expression in pre-B cells through ATM-FOXO1 signaling (29). They showed that the genotoxic drug NCS caused rapid downmodulation of Rag1 and Rag2 mRNA and Rag1 protein (they did not report on Rag2 protein) in STI571-treated human BCR-ABL + pre-B cell lines (BV173 and SUP-B15) and mouse v-Abl transformed pro-B cell lines.…”

Section: Discussionmentioning

confidence: 99%

“…As highlighted by our prior (24) and current Gadd45α data, a potential mechanistic link between release of FOXO1 and inhibition of Rag1/Rag2 transcription should be assessed to distinguish a cause-effect relationship versus another unlinked ATM-dependent correlation. Considering the complexities of ATM-FOXO1 signaling found by the Guikema lab (29), and the unresolved questions over the roles of Nemo and NF-κB proteins in Rag1/Rag2 transcription, major obstacles exist for testing these models. Further characterization of the molecular requirements and associated events of DSB-induced inhibition of Rag1/Rag2 expression should open paths forward.…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, the ability of ATM to signal nuclear export of a fraction of nuclear Rag2 protein in response to DSBs (46) opens the possibility that ATM might regulate the subcellular location of Rag1 protein. Finally, the finding of the Guikema lab (29) that the genotoxic drug neocarzinostatin increases the degradation of Rag1 protein suggests that ATM also might regulate the stability of Rag1. Clearly, studies are needed to investigate the potential role of ATM-dependent post-translational modifications of the Rag1 and Rag2 proteins in DSB-induced inhibition of V(D)J recombination.…”

Section: Discussionmentioning

confidence: 99%

“…In this study, we demonstrate that DSBs induced in primary lymphocytes by IR or etoposide signal through ATM and Nemo to transcriptionally repress Rag1 expression and inhibit RAG cleavage of Igκ loci and RSSs integrated at another genomic location. While we were preparing our manuscript, the Guikema lab reported that DSBs induced in immortalized and transformed human and mouse pre-B cell lines signal via ATM and the FOXO1 transcription factor to inhibit RAG expression and activity (29). We discuss how our study corroborates the central findings of the Guikema lab and presents novel data that provide further insights into the mechanisms and the scope by which lymphocytes direct DSB-induced feedback inhibition of V(D)J recombination.…”

Section: Introductionmentioning

confidence: 99%

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Immature Lymphocytes Inhibit Rag1 and Rag2 Transcription and V(D)J Recombination in Response to DNA Double-Strand Breaks

Fisher

Rivera-Reyes

Bloch

et al. 2017

The Journal of Immunology

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Mammalian cells have evolved a common DNA damage response (DDR) that sustains cellular function, maintains genomic integrity, and suppresses malignant transformation. In pre-B cells, DNA double strand breaks (DSBs) induced at Igκ loci by the Rag1/Rag2 (RAG) endonuclease engage this DDR to modulate transcription of genes that regulate lymphocyte-specific processes. We previously reported that RAG DSBs induced at one Igκ allele signal through the ATM kinase to feedback inhibit RAG expression and RAG cleavage of the other Igκ allele. Here, we show that DSBs induced by ionizing radiation, etoposide, or bleomycin suppress Rag1 and Rag2 mRNA levels in primary pre-B cells, pro-B cells, and pro-T cells, indicating that inhibition of Rag1 and Rag2 expression is a prevalent DSB response among immature lymphocytes. DSBs induced in pre-B cells signal rapid transcriptional repression of Rag1 and Rag2, causing downregulation of both Rag1 and Rag2 mRNA but only Rag1 protein. This transcriptional inhibition requires the ATM kinase and the NF-κB essential modulator protein, implicating a role for ATM-mediated activation of canonical NF-κB transcription factors. Finally, we demonstrate that DSBs induced in pre-B cells by etoposide or bleomycin inhibit recombination of Igκ loci and a chromosomally integrated substrate. Our data indicate that immature lymphocytes exploit a common DDR signaling pathway to limit DSBs at multiple genomic locations within developmental stages wherein monoallelic antigen receptor locus recombination is enforced. We discuss the implications of our findings for mechanisms that orchestrate the differentiation of mono-specific lymphocytes while suppressing oncogenic antigen receptor locus translocations.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Immature Lymphocytes Inhibit Rag1 and Rag2 Transcription and V(D)J Recombination in Response to DNA Double-Strand Breaks

Fisher

Rivera-Reyes

Bloch

et al. 2017

The Journal of Immunology

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Levamisole suppresses activation and proliferation of human T cells by the induction of a p53‐dependent DNA damage response

Khan,

Veltkamp,

Scheper

et al. 2023

Eur J Immunol

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Levamisole (LMS) is a small molecule used in the treatment of idiopathic nephrotic syndrome (INS). The pathogenesis of INS remains unknown, but evidence points towards an immunological basis of the disease. Recently, LMS has been shown to increase the relapse‐free survival in INS patients. While LMS has been hypothesized to exert an immunomodulatory effect, its mechanism of action remains unknown. Here, we show that LMS decreased activation and proliferation of human T cells. T‐cell activation‐associated cytokines such as IL‐2, TNF‐α and IFN‐γ were reduced upon LMS treatment, whereas IL‐4 and IL‐13 were increased. Gene expression profiling confirmed the suppressive effects of LMS as genes involved in cell cycle progression were downregulated. Furthermore, genes associated with p53 activation were upregulated by LMS. In agreement, LMS treatment resulted in p53 phosphorylation and increased expression of the p53 target gene FAS. Accordingly, LMS sensitized activated T cells for Fas‐mediated apoptosis. LMS treatment resulted in a mid‐S phase cell cycle arrest accompanied by γH2AX‐foci formation and phosphorylation of CHK1. Our findings indicate that LMS acts as an immunosuppressive drug that directly affects the activation and proliferation of human T cells by induction of DNA damage and the activation of a p53‐dependent DNA damage response.This article is protected by copyright. All rights reserved

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RAG1/2 induces double‐stranded DNA breaks at non‐Ig loci in the proximity of single sequence repeats in developing B cells

Ochodnicka‐Mackovicova,

Mokry,

Haagmans

et al. 2024

Eur J Immunol

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In developing B cells, V(D)J gene recombination is initiated by the RAG1/2 endonuclease complex, introducing double‐stranded DNA breaks (DSBs) in V, D, and J genes and resulting in the formation of the hypervariable parts of immunoglobulins (Ig). Persistent or aberrant RAG1/2 targeting is a potential threat to genome integrity. While RAG1 and RAG2 have been shown to bind various regions genome‐wide, the in vivo off‐target DNA damage instigated by RAG1/2 endonuclease remains less well understood. In the current study, we identified regions containing RAG1/2‐induced DNA breaks in mouse pre‐B cells on a genome‐wide scale using a global DNA DSB detection strategy. We detected 1489 putative RAG1/2‐dependent DSBs, most of which were located outside the Ig loci. DNA sequence motif analysis showed a specific enrichment of RAG1/2‐induced DNA DSBs at GA‐ and CA‐repeats and GC‐rich motifs. These findings provide further insights into RAG1/2 off‐target activity. The ability of RAG1/2 to introduce DSBs on the non‐Ig loci during the endogenous V(D)J recombination emphasizes its genotoxic potential in developing lymphocytes.

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The DNA Damage Response Regulates RAG1/2 Expression in Pre–B Cells through ATM-FOXO1 Signaling

Cited by 30 publications

References 62 publications

Immature Lymphocytes Inhibit Rag1 and Rag2 Transcription and V(D)J Recombination in Response to DNA Double-Strand Breaks

Immature Lymphocytes Inhibit Rag1 and Rag2 Transcription and V(D)J Recombination in Response to DNA Double-Strand Breaks

Levamisole suppresses activation and proliferation of human T cells by the induction of a p53‐dependent DNA damage response

RAG1/2 induces double‐stranded DNA breaks at non‐Ig loci in the proximity of single sequence repeats in developing B cells

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