the brct domain of parp-1 is required for immunoglobulin gene conversionbrct域所需的parp-1是免疫球蛋白基因转换.pdfVIP

The BRCT Domain of PARP-1 Is Required for Immunoglobulin Gene Conversion 1,2 1,2 3 1,2 Marcia N. Paddock , Ben D. Buelow , Shunichi Takeda , Andrew M. Scharenberg * 1 Department of Immunology, University of Washington, Seattle, Washington, United States of America, 2 Center for Immunity and Immunotherapies, Seattle Children’s Hospital Research Institute, Seattle, Washington, United States of America, 3 Crest Laboratory, Department of Radiation Genetics, Faculty of Medicine, Kyoto University, Sakyo-ku, Kyoto, Japan Abstract Genetic variation at immunoglobulin (Ig) gene variable regions in B-cells is created through a multi-step process involving deamination of cytosine bases by activation-induced cytidine deaminase (AID) and their subsequent mutagenic repair. To protect the genome from dangerous, potentially oncogenic effects of off-target mutations, both AID activity and mutagenic repair are targeted specifically to the Ig genes. However, the mechanisms of targeting are unknown and recent data have highlighted the role of regulating mutagenic repair to limit the accumulation of somatic mutations resulting from the more widely distributed AID-induced lesions to the Ig genes. Here we investigated the role of the DNA damage sensor poly- (ADPribose)-polymerase-1 (PARP-1) in the repair of AID-induced DNA lesions. We show through sequencing of the diversifying Ig genes in PARP-12/ 2 DT40 B-cells that PARP-1 deficiency results in a marked reduction in gene conversion events and enhanced high-fidelity repair of AID-induced lesions at both Ig heavy and light chains. To further characterize the role of PARP-1 in the mutagenic repair of AID-induced lesions, we