Format

Send to

Choose Destination
Nat Commun. 2018 May 22;9(1):2016. doi: 10.1038/s41467-018-04466-4.

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

Author information

1
The CRUK Gene Function Laboratory and Breast Cancer Now Toby Robins Research Centre, The Institute of Cancer Research, London, SW3 6JB, UK.
2
The Cancer Therapeutics Division, The Institute of Cancer Research, London, SW3 6JB, UK.
3
The Institute of Molecular Biology, Bulgarian Academy of Sciences, 1113, Sofia, Bulgaria.
4
The CRUK Gene Function Laboratory and Breast Cancer Now Toby Robins Research Centre, The Institute of Cancer Research, London, SW3 6JB, UK. Alan.Ashworth@ucsf.edu.
5
UCSF Helen Diller Family Comprehensive Cancer Center, 1450 3rd Street, San Francisco, CA, 94158, USA. Alan.Ashworth@ucsf.edu.
6
The CRUK Gene Function Laboratory and Breast Cancer Now Toby Robins Research Centre, The Institute of Cancer Research, London, SW3 6JB, UK. Chris.Lord@icr.ac.uk.

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.

PMID:
29789535
PMCID:
PMC5964205
DOI:
10.1038/s41467-018-04466-4
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center