Localized protein biotinylation at DNA damage sites identifies ZPET, a repressor of homologous recombination

Numerous DNA repair and signaling proteins function at DNA damage sites to protect the genome. Here, we show that fusion of the promiscuous biotin ligase BirA with RAD18 leads to localized protein biotinylation at DNA damage sites, allowing identification of ZPET (zinc finger protein proximal to RAD...

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Bibliographic Details
Published in:Genes & development 2019-01, Vol.33 (1-2), p.75-89
Main Authors: Moquin, David M, Genois, Marie-Michelle, Zhang, Jia-Min, Ouyang, Jian, Yadav, Tribhuwan, Buisson, Rémi, Yazinski, Stephanie A, Tan, Jun, Boukhali, Myriam, Gagné, Jean-Philippe, Poirier, Guy G, Lan, Li, Haas, Wilhelm, Zou, Lee
Format: Article
Language:English
Subjects:
Biotinylation - methods
Carbon-Nitrogen Ligases - genetics
Cell Line
DNA Breaks, Double-Stranded
DNA Damage
DNA Repair - genetics
DNA, Single-Stranded - metabolism
DNA-Binding Proteins - genetics
DNA-Binding Proteins - metabolism
Escherichia coli Proteins - genetics
Gene Knockdown Techniques
Homologous Recombination - genetics
Humans
MRE11 Homologue Protein - metabolism
Protein Binding
Protein Transport - genetics
Repressor Proteins - genetics
Research Paper
Transcription Factors - genetics
Transcription Factors - metabolism
Ubiquitin-Protein Ligases - genetics
Ubiquitin-Protein Ligases - metabolism
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Summary:Numerous DNA repair and signaling proteins function at DNA damage sites to protect the genome. Here, we show that fusion of the promiscuous biotin ligase BirA with RAD18 leads to localized protein biotinylation at DNA damage sites, allowing identification of ZPET (zinc finger protein proximal to RAD eighteen)/ZNF280C as a potential DNA damage response (DDR) protein. ZPET binds ssDNA and localizes to DNA double-strand breaks (DSBs) and stalled replication forks. In vitro, ZPET inhibits MRE11 binding to ssDNA. In cells, ZPET delays MRE11 binding to chromatin after DSB formation and slows DNA end resection through binding ssDNA. ZPET hinders resection independently of 53BP1 and HELB. Cells lacking ZPET displayed enhanced homologous recombination (HR), accelerated replication forks under stress, and increased resistance to DSBs and PARP inhibition. These results not only reveal ZPET as an HR repressor but also suggest that localized protein biotinylation at DNA damage sites is a useful strategy to identify DDR proteins.
ISSN:0890-9369
1549-5477
DOI:10.1101/gad.315978.118