Coupling bimolecular PARylation biosensors with genetic screens to identify PARylation targets.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 29789535.
- Also identified by DOI 10.1038/s41467-018-04466-4 and PMC identifier 5964205.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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.
Medical subject headings
- Biosensing Techniques
- Eukaryotic Initiation Factors
- Mitosis
- Protein Processing, Post-Translational
- Tankyrases