A Damage-Independent Role for 53BP1 that Impacts Break Order and Igh Architecture during Class Switch Recombination

Pedro P. Rocha, Ramya Raviram, Yi Fu, Jung Hyun Kim, Vincent M. Luo, Arafat Aljoufi, Emily Swanzey, Alessandra Pasquarella, Alessia Balestrini, Emily R. Miraldi, Richard Bonneau, John Petrini, Gunnar Schotta, Jane A. Skok

Research output: Contribution to journalArticle

Abstract

During class switch recombination (CSR), B cells replace the Igh Cμ or δ exons with another downstream constant region exon (CH), altering the antibody isotype. CSR occurs through the introduction of AID-mediated double-strand breaks (DSBs) in switch regions and subsequent ligation of broken ends. Here, we developed an assay to investigate the dynamics of DSB formation in individual cells. We demonstrate that the upstream switch region Sμ is first targeted during recombination and that the mechanism underlying this control relies on 53BP1. Surprisingly, regulation of break order occurs through residual binding of 53BP1 to chromatin before the introduction of damage and independent of its established role in DNA repair. Using chromosome conformation capture, we show that 53BP1 mediates changes in chromatin architecture that affect break order. Finally, our results explain how changes in Igh architecture in the absence of 53BP1 could promote inversional rearrangements that compromise CSR. Rocha et al. describe a role for the DNA-damage sensor 53BP1 that is independent of the DNA double-strand break repair pathway. Residual recruitment of 53BP1 to chromatin impacts the 3D structure of . Igh during CSR and is necessary for ordered AID-mediated break formation on switch regions.

Original languageEnglish (US)
JournalCell Reports
DOIs
StateAccepted/In press - Feb 23 2016

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Genetic Recombination
Switches
Chromatin
Exons
DNA
Double-Stranded DNA Breaks
Repair
DNA Repair
DNA Damage
Ligation
B-Lymphocytes
Chromosomes
Conformations
Assays
Antibodies
Cells
Sensors

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

A Damage-Independent Role for 53BP1 that Impacts Break Order and Igh Architecture during Class Switch Recombination. / Rocha, Pedro P.; Raviram, Ramya; Fu, Yi; Kim, Jung Hyun; Luo, Vincent M.; Aljoufi, Arafat; Swanzey, Emily; Pasquarella, Alessandra; Balestrini, Alessia; Miraldi, Emily R.; Bonneau, Richard; Petrini, John; Schotta, Gunnar; Skok, Jane A.

In: Cell Reports, 23.02.2016.

Research output: Contribution to journalArticle

Rocha, PP, Raviram, R, Fu, Y, Kim, JH, Luo, VM, Aljoufi, A, Swanzey, E, Pasquarella, A, Balestrini, A, Miraldi, ER, Bonneau, R, Petrini, J, Schotta, G & Skok, JA 2016, 'A Damage-Independent Role for 53BP1 that Impacts Break Order and Igh Architecture during Class Switch Recombination', Cell Reports. https://doi.org/10.1016/j.celrep.2016.05.073
Rocha, Pedro P. ; Raviram, Ramya ; Fu, Yi ; Kim, Jung Hyun ; Luo, Vincent M. ; Aljoufi, Arafat ; Swanzey, Emily ; Pasquarella, Alessandra ; Balestrini, Alessia ; Miraldi, Emily R. ; Bonneau, Richard ; Petrini, John ; Schotta, Gunnar ; Skok, Jane A. / A Damage-Independent Role for 53BP1 that Impacts Break Order and Igh Architecture during Class Switch Recombination. In: Cell Reports. 2016.
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