Exo1 protein, mouse; Exodeoxyribonucleases; DNA Repair Enzymes; Animals; B-Lymphocytes; Immunity; Meiosis/genetics; Mice; Somatic Hypermutation, Immunoglobulin; DNA Repair; DNA Repair Enzymes/genetics; DNA Repair Enzymes/metabolism; Exodeoxyribonucleases/genetics; Exodeoxyribonucleases/metabolism; Neoplasms/genetics; Neoplasms/immunology; Genetics
Abstract :
[en] DNA damage response pathways rely extensively on nuclease activity to process DNA intermediates. Exonuclease 1 (EXO1) is a pleiotropic evolutionary conserved DNA exonuclease involved in various DNA repair pathways, replication, antibody diversification, and meiosis. But, whether EXO1 facilitates these DNA metabolic processes through its enzymatic or scaffolding functions remains unclear. Here, we dissect the contribution of EXO1 enzymatic versus scaffolding activity by comparing Exo1DA/DA mice expressing a proven nuclease-dead mutant form of EXO1 to entirely EXO1-deficient Exo1-/- and EXO1 wild type Exo1+/+ mice. We show that Exo1DA/DA and Exo1-/- mice are compromised in canonical DNA repair processing, suggesting that the EXO1 enzymatic role is important for error-free DNA mismatch and double-strand break repair pathways. However, in non-canonical repair pathways, EXO1 appears to have a more nuanced function. Next-generation sequencing of heavy chain V region in B cells showed the mutation spectra of Exo1DA/DA mice to be intermediate between Exo1+/+ and Exo1-/- mice, suggesting that both catalytic and scaffolding roles of EXO1 are important for somatic hypermutation. Similarly, while overall class switch recombination in Exo1DA/DA and Exo1-/- mice was comparably defective, switch junction analysis suggests that EXO1 might fulfill an additional scaffolding function downstream of class switching. In contrast to Exo1-/- mice that are infertile, meiosis progressed normally in Exo1DA/DA and Exo1+/+ cohorts, indicating that a structural but not the nuclease function of EXO1 is critical for meiosis. However, both Exo1DA/DA and Exo1-/- mice displayed similar mortality and cancer predisposition profiles. Taken together, these data demonstrate that EXO1 has both scaffolding and enzymatic functions in distinct DNA repair processes and suggest a more composite and intricate role for EXO1 in DNA metabolic processes and disease.
Disciplines :
Genetics & genetic processes
Author, co-author :
Wang, Shanzhi; Department of Cell Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, NY 10461, USA ; Current position: Department of Chemistry, University of Arkansas at Little Rock, Little Rock, AR 72204, USA
Lee, Kyeryoung; Department of Cell Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, NY 10461, USA
Gray, Stephen ; Department of Biomedical Sciences, Cornell University, NY 14853, USA ; Current position: School of Life Sciences, University of Nottingham, Nottingham NG7 2UH, UK
Zhang, Yongwei; Department of Cell Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, NY 10461, USA
Tang, Catherine; Department of Applied Mathematics and Statistics, Stony Brook University, Stony Brook, NY, USA
Morrish, Rikke B; Current position: School of Physics and Astronomy, University of Exeter, Exeter EX4 4QD, UK
Tosti, Elena; Department of Cell Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, NY 10461, USA
van Oers, Johanna; Department of Cell Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, NY 10461, USA
Amin, Mohammad Ruhul; Department of Computer and Information Science, Fordham University, Bronx, NY, USA
Cohen, Paula E; Department of Biomedical Sciences, Cornell University, NY 14853, USA
MacCarthy, Thomas; Department of Applied Mathematics and Statistics, Stony Brook University, Stony Brook, NY, USA
Roa, Sergio ; Department of Biochemistry and Genetics, University of Navarra, 31008Pamplona, Spain ; Centro de Investigacion Biomedica en Red de Cancer (CIBERONC), Instituto de Salud Carlos III, Madrid, Spain ; Navarra Institute for Health Research (IdiSNA), Pamplona, Spain
Scharff, Matthew D; Department of Cell Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, NY 10461, USA
Edelmann, Winfried; Department of Cell Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, NY 10461, USA
CHAHWAN, Richard ; University of Luxembourg ; Institute of Experimental Immunology, University of Zurich, Zurich 8057, Switzerland
NIH NIH NIH NIH NIH NIH BBSRC SNF Vontobel Agencia Estatal de Investigación Ministerio de Economía, Industria y Competitividad Harry Eagle Chair Ministerio de Economía y Competitividad AMS EPSRC
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