[en] An effective immune response requires B cells to produce several classes of antibodies through the process of class switch recombination (CSR). Activation-induced cytidine deaminase initiates CSR by deaminating deoxycytidines at switch regions within the Ig locus. This activity leads to double-stranded DNA break formation at the donor and recipient switch regions that are subsequently synapsed and ligated in a 53BP1-dependent process that remains poorly understood. The DNA damage response E3 ubiquitin ligases RNF8 and RNF168 were recently shown to facilitate recruitment of 53BP1 to sites of DNA damage. Here we show that the ubiquitination pathway mediated by RNF8 and RNF168 plays an integral part in CSR. Using the CH12F3-2 mouse B cell line that undergoes CSR to IgA at high rates, we demonstrate that knockdown of RNF8, RNF168, and 53BP1 leads to a significant decrease in CSR. We also show that 53BP1-deficient CH12F3-2 cells are protected from apoptosis mediated by the MDM2 inhibitor Nutlin-3. In contrast, deficiency in either E3 ubiquitin ligase does not protect cells from Nutlin-3-mediated apoptosis, indicating that RNF8 and RNF168 do not regulate all functions of 53BP1.
Disciplines :
Immunology & infectious disease
Author, co-author :
Ramachandran, Shaliny; Department of Immunology, University of Toronto, Toronto, Ontario, Canada
CHAHWAN, Richard ; University of Luxembourg > Faculty of Science, Technology and Medicine (FSTM) > Department of Health, Medicine and Life Sciences (DHML) ; Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY, United States
Nepal, Rajeev M; Department of Immunology, University of Toronto, Toronto, ON, Canada
Frieder, Darina; Department of Immunology, University of Toronto, Toronto, ON, Canada
Panier, Stephanie; Samuel Lunenfeld Research Institute, Mount Sinai Hospital and Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada
Roa, Sergio; Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY, United States
Zaheen, Ahmad; Department of Immunology, University of Toronto, Toronto, ON, Canada
Durocher, Daniel; Samuel Lunenfeld Research Institute, Mount Sinai Hospital and Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada
Scharff, Matthew D; Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY, United States
Martin, Alberto; Department of Immunology, University of Toronto, Toronto, ON, Canada
External co-authors :
yes
Language :
English
Title :
The RNF8/RNF168 ubiquitin ligase cascade facilitates class switch recombination.
Publication date :
12 January 2010
Journal title :
Proceedings of the National Academy of Sciences of the United States of America
ISSN :
0027-8424
eISSN :
1091-6490
Publisher :
Proceedings of the National Academy of Sciences, United States
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