Cargando…
Coupling bimolecular PARylation biosensors with genetic screens to identify PARylation targets
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 linkin...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5964205/ https://www.ncbi.nlm.nih.gov/pubmed/29789535 http://dx.doi.org/10.1038/s41467-018-04466-4 |
_version_ | 1783325138833375232 |
---|---|
author | Krastev, Dragomir B. Pettitt, Stephen J. Campbell, James Song, Feifei Tanos, Barbara E. Stoynov, Stoyno S. Ashworth, Alan Lord, Christopher J. |
author_facet | Krastev, Dragomir B. Pettitt, Stephen J. Campbell, James Song, Feifei Tanos, Barbara E. Stoynov, Stoyno S. Ashworth, Alan Lord, Christopher J. |
author_sort | Krastev, Dragomir B. |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-5964205 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-59642052018-05-24 Coupling bimolecular PARylation biosensors with genetic screens to identify PARylation targets Krastev, Dragomir B. Pettitt, Stephen J. Campbell, James Song, Feifei Tanos, Barbara E. Stoynov, Stoyno S. Ashworth, Alan Lord, Christopher J. Nat Commun Article 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. Nature Publishing Group UK 2018-05-22 /pmc/articles/PMC5964205/ /pubmed/29789535 http://dx.doi.org/10.1038/s41467-018-04466-4 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Krastev, Dragomir B. Pettitt, Stephen J. Campbell, James Song, Feifei Tanos, Barbara E. Stoynov, Stoyno S. Ashworth, Alan Lord, Christopher J. Coupling bimolecular PARylation biosensors with genetic screens to identify PARylation targets |
title | Coupling bimolecular PARylation biosensors with genetic screens to identify PARylation targets |
title_full | Coupling bimolecular PARylation biosensors with genetic screens to identify PARylation targets |
title_fullStr | Coupling bimolecular PARylation biosensors with genetic screens to identify PARylation targets |
title_full_unstemmed | Coupling bimolecular PARylation biosensors with genetic screens to identify PARylation targets |
title_short | Coupling bimolecular PARylation biosensors with genetic screens to identify PARylation targets |
title_sort | coupling bimolecular parylation biosensors with genetic screens to identify parylation targets |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5964205/ https://www.ncbi.nlm.nih.gov/pubmed/29789535 http://dx.doi.org/10.1038/s41467-018-04466-4 |
work_keys_str_mv | AT krastevdragomirb couplingbimolecularparylationbiosensorswithgeneticscreenstoidentifyparylationtargets AT pettittstephenj couplingbimolecularparylationbiosensorswithgeneticscreenstoidentifyparylationtargets AT campbelljames couplingbimolecularparylationbiosensorswithgeneticscreenstoidentifyparylationtargets AT songfeifei couplingbimolecularparylationbiosensorswithgeneticscreenstoidentifyparylationtargets AT tanosbarbarae couplingbimolecularparylationbiosensorswithgeneticscreenstoidentifyparylationtargets AT stoynovstoynos couplingbimolecularparylationbiosensorswithgeneticscreenstoidentifyparylationtargets AT ashworthalan couplingbimolecularparylationbiosensorswithgeneticscreenstoidentifyparylationtargets AT lordchristopherj couplingbimolecularparylationbiosensorswithgeneticscreenstoidentifyparylationtargets |