title: RAG-mediated DNA double-strand breaks activate a cell type-specific checkpoint to inhibit pre-B cell receptor signals.
abstract: DNA double-strand breaks (DSBs) activate a canonical DNA damage response, including highly conserved cell cycle checkpoint pathways that prevent cells with DSBs from progressing through the cell cycle. In developing B cells, pre-B cell receptor (pre-BCR) signals initiate immunoglobulin light (Igl) chain gene assembly, leading to RAG-mediated DNA DSBs. The pre-BCR also promotes cell cycle entry, which could cause aberrant DSB repair and genome instability in pre-B cells. Here, we show that RAG DSBs inhibit pre-BCR signals through the ATM- and NF-κB2-dependent induction of SPIC, a hematopoietic-specific transcriptional repressor. SPIC inhibits expression of the SYK tyrosine kinase and BLNK adaptor, resulting in suppression of pre-BCR signaling. This regulatory circuit prevents the pre-BCR from inducing additional Igl chain gene rearrangements and driving pre-B cells with RAG DSBs into cycle. We propose that pre-B cells toggle between pre-BCR signals and a RAG DSB-dependent checkpoint to maintain genome stability while iteratively assembling Igl chain genes.
authors.affiliation: Department of Pediatrics, Washington University School of Medicine, St. Louis, MO 63110.
authors.forename: Jeffrey J
authors.initials: JJ
authors.lastname: Bednarski
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110.
authors.forename: Ruchi
authors.initials: R
authors.lastname: Pandey
authors.affiliation: Department of Pediatrics, Washington University School of Medicine, St. Louis, MO 63110.
authors.forename: Emily
authors.initials: E
authors.lastname: Schulte
authors.affiliation: Department of Pediatrics, Washington University School of Medicine, St. Louis, MO 63110.
authors.forename: Lynn S
authors.initials: LS
authors.lastname: White
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110.
authors.forename: Bo-Ruei
authors.initials: BR
authors.lastname: Chen
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110.
authors.forename: Gabriel J
authors.initials: GJ
authors.lastname: Sandoval
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110.
authors.forename: Masako
authors.initials: M
authors.lastname: Kohyama
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110.
authors.forename: Malay
authors.initials: M
authors.lastname: Haldar
authors.affiliation: Department of Pediatrics, Washington University School of Medicine, St. Louis, MO 63110.
authors.forename: Andrew
authors.initials: A
authors.lastname: Nickless
authors.affiliation: Department of Pediatrics, Washington University School of Medicine, St. Louis, MO 63110.
authors.forename: Amanda
authors.initials: A
authors.lastname: Trott
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, CA 90095.
authors.forename: Genhong
authors.initials: G
authors.lastname: Cheng
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110.
authors.forename: Kenneth M
authors.initials: KM
authors.lastname: Murphy
authors.affiliation: Division of Cancer Pathobiology, Department of Pathology and Laboratory Medicine, Center for Childhood Cancer Research, Children's Hospital of Philadelphia, Philadelphia, PA 19104.
authors.forename: Craig H
authors.initials: CH
authors.lastname: Bassing
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110.
authors.forename: Jacqueline E
authors.initials: JE
authors.lastname: Payton
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110 bas2022@med.cornell.edu.
authors.forename: Barry P
authors.initials: BP
authors.lastname: Sleckman
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000818:Animals; D064007:Ataxia Telangiectasia Mutated Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D059447:Cell Cycle Checkpoints / Q000276:immunology; D049109:Cell Proliferation; D053903:DNA Breaks, Double-Stranded; D004268:DNA-Binding Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D015328:Gene Rearrangement, B-Lymphocyte, Light Chain; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D052003:NF-kappa B p52 Subunit / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D054420:Pre-B Cell Receptors / Q000378:metabolism*; D054448:Precursor Cells, B-Lymphoid / Q000166:cytology / Q000276:immunology* / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D015398:Signal Transduction / Q000276:immunology; D000072377:Syk Kinase; D015534:Trans-Activators / Q000378:metabolism; D000097782:NF-kappaB-Inducing Kinase; D000099232:Proto-Oncogene Protein Spi-1
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D064007:Ataxia Telangiectasia Mutated Proteins; D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D047908:Intracellular Signaling Peptides and Proteins; D052003:NF-kappa B p52 Subunit; D054420:Pre-B Cell Receptors; D017346:Protein Serine-Threonine Kinases; D011505:Protein-Tyrosine Kinases; D011518:Proto-Oncogene Proteins; D000072377:Syk Kinase; D015534:Trans-Activators; D000097782:NF-kappaB-Inducing Kinase; D000099232:Proto-Oncogene Protein Spi-1; C113939:B cell linker protein; C498207:Nfkb2 protein, mouse; C423918:Spic protein, mouse; C064658:RAG-1 protein; C000606897:Syk protein, mouse; C576325:Atm protein, mouse
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