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Coupling bimolecular PARylation biosensors with genetic screens to identify PARylation targets

Author

Listed:
  • Dragomir B. Krastev

    (The Institute of Cancer Research)

  • Stephen J. Pettitt

    (The Institute of Cancer Research)

  • James Campbell

    (The Institute of Cancer Research)

  • Feifei Song

    (The Institute of Cancer Research)

  • Barbara E. Tanos

    (The Institute of Cancer Research)

  • Stoyno S. Stoynov

    (Bulgarian Academy of Sciences)

  • Alan Ashworth

    (The Institute of Cancer Research
    UCSF Helen Diller Family Comprehensive Cancer Center)

  • Christopher J. Lord

    (The Institute of Cancer Research)

Abstract

Poly (ADP-ribose)ylation is a dynamic protein modification that regulates multiple cellular processes. Here, we describe a system for identifying and characterizing PARylation events that exploits the ability of a PBZ (PAR-binding zinc finger) protein domain to bind PAR with high-affinity. By linking PBZ domains to bimolecular fluorescent complementation biosensors, we developed fluorescent PAR biosensors that allow the detection of temporal and spatial PARylation events in live cells. Exploiting transposon-mediated recombination, we integrate the PAR biosensor en masse into thousands of protein coding genes in living cells. Using these PAR-biosensor “tagged” cells in a genetic screen we carry out a large-scale identification of PARylation targets. This identifies CTIF (CBP80/CBP20-dependent translation initiation factor) as a novel PARylation target of the tankyrase enzymes in the centrosomal region of cells, which plays a role in the distribution of the centrosomal satellites.

Suggested Citation

  • Dragomir B. Krastev & Stephen J. Pettitt & James Campbell & Feifei Song & Barbara E. Tanos & Stoyno S. Stoynov & Alan Ashworth & Christopher J. Lord, 2018. "Coupling bimolecular PARylation biosensors with genetic screens to identify PARylation targets," Nature Communications, Nature, vol. 9(1), pages 1-14, December.
    • Handle: RePEc:nat:natcom:v:9:y:2018:i:1:d:10.1038_s41467-018-04466-4
    DOI: 10.1038/s41467-018-04466-4
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