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Shieldin and CST co-orchestrate DNA polymerase-dependent tailed-end joining reactions independently of 53BP1-governed repair pathway choice

Abstract:
Tumor suppressor p53-binding protein 1 (53BP1) regulates DNA end joining in lymphocytes, diversifying immune antigen receptors. This involves nucleosome-bound 53BP1 at DNA double-stranded breaks (DSBs) recruiting Rap1-interacting factor 1 homolog (RIF1) and shieldin, a poorly understood DNA-binding complex. The 53BP1–RIF1–shieldin axis is pathological in BRCA1-mutated cancers, blocking homologous recombination (HR) and driving illegitimate nonhomologous end joining (NHEJ). However, how this axis regulates DNA end joining and HR suppression remains unresolved. We investigated shieldin and its interplay with the Ctc1–Stn1–Ten1 (CST) complex, which was recently implicated downstream of 53BP1. Immunophenotypically, mice lacking shieldin or CST are equivalent, with class-switch recombination coreliant on both complexes. Ataxia-telangiectasia mutated kinase-dependent DNA damage signaling underpins this cooperation, inducing physical interactions between these complexes that reveal shieldin as a DSB-responsive CST adaptor. Furthermore, DNA polymerase ζ functions downstream of shieldin, establishing DNA fill-in synthesis as the physiological function of shieldin–CST. Lastly, we demonstrate that 53BP1 suppresses HR and promotes NHEJ in BRCA1-deficient mice and cells independently of shieldin. These findings showcase the versatility of the 53BP1 pathway, achieved through the collaboration of chromatin-bound 53BP1 complexes and DNA end-processing effector proteins.
Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1038/s41594-024-01381-9

Authors


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Institution:
University of Oxford
Division:
MSD
Department:
RDM
Sub department:
Weatherall Insti. of Molecular Medicine
Research group:
Genome Integrity laboratory
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MSD
Department:
RDM
Sub department:
RDM Clinical Laboratory Sciences
Research group:
Genome Integrity laboratory
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MSD
Department:
RDM
Sub department:
Weatherall Insti. of Molecular Medicine
Research group:
Genome Integrity laboratory
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MSD
Department:
NDM
Research group:
Centre for ImmunoOncology
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MSD
Department:
RDM
Sub department:
Weatherall Insti. of Molecular Medicine
Research group:
Genome Integrity laboratory
Role:
Author


More from this funder
Funder identifier:
https://ror.org/054225q67
Grant:
RCCSCF-Nov21\100004
19270
More from this funder
Funder identifier:
https://ror.org/03x94j517
Grant:
MR/R017549/1
MR/M009971/1
More from this funder
Funder identifier:
https://ror.org/03356n642


Publisher:
Springer Nature
Journal:
Nature Structural and Molecular Biology More from this journal
Volume:
32
Issue:
1
Pages:
86–97
Publication date:
2024-09-03
Acceptance date:
2024-08-01
DOI:
EISSN:
1545-9985
ISSN:
1545-9993


Language:
English
Pubs id:
2022024
Local pid:
pubs:2022024
Deposit date:
2024-08-16

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