title: DNA-dependent protein kinase mediates V(D)J recombination via RAG2 phosphorylation.
abstract: V(D)J recombination, a site-specific gene rearrangement process occurring during the lymphocyte development, begins with DNA double strand breaks by two recombination activating gene products (RAG1/2) and finishes with the repair process by several proteins including DNA-dependent protein kinase (DNA-PK). In this report, we found that RAG2 was specifically phosphorylated by DNA-PK at the 365(th) serine residue, and this phosphorylated RAG2 affected the V(D)J recombination activity in cells in the GFP expression-based assay. While the V(D)J recombination activity between wild-type RAG2 and mutant S365A RAG2 in the assay using a signal joint substrate was undistinguishable in DNA-PK deficient cells (M059J), the activity with wild-type RAG2 was largely increased in DNA-PK proficient cells (M059K) in comparison with mutant RAG2, suggesting that RAG2 phosphorylation by DNA-PK plays a crucial role in the signal joint formation during V(D)J recombination.
authors.affiliation: Department of Biochemistry and MRCND, School of Medicine and Gyeongsang Institute of Health Sciences, Gyeongsang National University, JinJu, Republic of Korea.
authors.forename: Young-Sool
authors.initials: YS
authors.lastname: Hah
authors.forename: Jung Hwa
authors.initials: JH
authors.lastname: Lee
authors.forename: Deok Ryong
authors.initials: DR
authors.lastname: Kim
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D045744:Cell Line, Tumor; D051747:DNA-Activated Protein Kinase / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D015321:Gene Rearrangement; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D016415:Sequence Alignment; D014162:Transfection; D045567:VDJ Recombinases / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; C499311:RAG2 protein, human; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D051747:DNA-Activated Protein Kinase; D045567:VDJ Recombinases