title: Spatio-temporal regulation of RAG2 following genotoxic stress.
abstract: V(D)J recombination of lymphocyte antigen receptor genes occurs via the formation of DNA double strand breaks (DSBs) through the activity of RAG1 and RAG2. The co-existence of RAG-independent DNA DSBs generated by genotoxic stressors potentially increases the risk of incorrect repair and chromosomal abnormalities. However, it is not known whether cellular responses to DSBs by genotoxic stressors affect the RAG complex. Using cellular imaging and subcellular fractionation approaches, we show that formation of DSBs by treating cells with DNA damaging agents causes export of nuclear RAG2. Within the cytoplasm, RAG2 exhibited substantial enrichment at the centrosome. Further, RAG2 export was sensitive to inhibition of ATM, and was reversed following DNA repair. The core region of RAG2 was sufficient for export, but not centrosome targeting, and RAG2 export was blocked by mutation of Thr(490). In summary, DNA damage triggers relocalization of RAG2 from the nucleus to centrosomes, suggesting a novel mechanism for modulating cellular responses to DSBs in developing lymphocytes.
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA; Department of Microbiology and Immunology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA; Department of Pathology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.
authors.forename: William
authors.initials: W
authors.lastname: Rodgers
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.
authors.forename: Jennifer N
authors.initials: JN
authors.lastname: Byrum
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.
authors.forename: Hem
authors.initials: H
authors.lastname: Sapkota
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.
authors.forename: Negar S
authors.initials: NS
authors.lastname: Rahman
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.
authors.forename: Robert C
authors.initials: RC
authors.lastname: Cail
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.
authors.forename: Shuying
authors.initials: S
authors.lastname: Zhao
authors.affiliation: Department of Immunobiology and Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, CT, USA.
authors.forename: David G
authors.initials: DG
authors.lastname: Schatz
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA. Electronic address: karla-rodgers@ouhsc.edu.
authors.forename: Karla K
authors.initials: KK
authors.lastname: Rodgers
mesh_terms: D021581:Active Transport, Cell Nucleus; D064007:Ataxia Telangiectasia Mutated Proteins / Q000378:metabolism / Q000502:physiology; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D018385:Centrosome / Q000378:metabolism*; D004247:DNA / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D006801:Humans; D008856:Microscopy, Fluorescence; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D054448:Precursor Cells, B-Lymphoid / Q000378:metabolism*; D011839:Radiation, Ionizing; D013347:Subcellular Fractions / Q000378:metabolism; D045567:VDJ Recombinases / Q000235:genetics / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; C499311:RAG2 protein, human; D004247:DNA; C576324:ATM protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; D045567:VDJ Recombinases
keywords: Centrosome; DNA damage response; Genotoxic stress; RAG1; RAG2; V(D)J recombination