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Coupling bimolecular PARylation biosensors with genetic screens to identify PARylation targets.


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.

SUBMITTER: Krastev DB 

PROVIDER: S-EPMC5964205 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature

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Coupling bimolecular PARylation biosensors with genetic screens to identify PARylation targets.

Krastev Dragomir B DB   Pettitt Stephen J SJ   Campbell James J   Song Feifei F   Tanos Barbara E BE   Stoynov Stoyno S SS   Ashworth Alan A   Lord Christopher J CJ  

Nature communications 20180522 1


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. Exploitin  ...[more]

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