Coupling bimolecular PARylation biosensors with genetic screens to identify PARylation targets.
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Date
2018-05-22Author
Krastev, DB
Pettitt, SJ
Campbell, J
Song, F
Tanos, BE
Stoynov, SS
Ashworth, A
Lord, CJ
Type
Journal Article
Metadata
Show full item recordAbstract
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.
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Subject
Cell Line, Tumor
Hela Cells
Centrosome
Centrioles
Epithelial Cells
Humans
Tankyrases
Poly Adenosine Diphosphate Ribose
Calcium-Binding Proteins
Eukaryotic Initiation Factors
DNA Transposable Elements
Fluorescent Dyes
Biosensing Techniques
Signal Transduction
Mitosis
Protein Processing, Post-Translational
Recombination, Genetic
Genetic Testing
Poly ADP Ribosylation
Research team
Gene Function
Language
eng
Date accepted
2018-05-01
License start date
2018-05-22
Citation
Nature communications, 2018, 9 (1), pp. 2016 - ?
Publisher
NATURE PUBLISHING GROUP