Genetically Encoded Fluorescent Sensor for Poly-ADP-Ribose.
Bacterial Proteins
/ analysis
Biosensing Techniques
/ methods
Cell Line
Fluorescence Resonance Energy Transfer
/ methods
Fluorescent Dyes
/ analysis
Humans
Luminescent Proteins
/ analysis
Open Reading Frames
Poly Adenosine Diphosphate Ribose
/ analysis
Protein Domains
Recombinant Fusion Proteins
/ analysis
Ubiquitin-Protein Ligases
/ analysis
DNA damage respose
FRET
PAR
WWE-domai
fluorescent protein
sensor
Journal
International journal of molecular sciences
ISSN: 1422-0067
Titre abrégé: Int J Mol Sci
Pays: Switzerland
ID NLM: 101092791
Informations de publication
Date de publication:
15 Jul 2020
15 Jul 2020
Historique:
received:
12
06
2020
revised:
06
07
2020
accepted:
13
07
2020
entrez:
19
7
2020
pubmed:
19
7
2020
medline:
20
2
2021
Statut:
epublish
Résumé
Poly-(ADP-ribosyl)-ation (PARylation) is a reversible post-translational modification of proteins and DNA that plays an important role in various cellular processes such as DNA damage response, replication, transcription, and cell death. Here we designed a fully genetically encoded fluorescent sensor for poly-(ADP-ribose) (PAR) based on Förster resonance energy transfer (FRET). The WWE domain, which recognizes iso-ADP-ribose internal PAR-specific structural unit, was used as a PAR-targeting module. The sensor consisted of cyan Turquoise2 and yellow Venus fluorescent proteins, each in fusion with the WWE domain of RNF146 E3 ubiquitin ligase protein. This bipartite sensor named sPARroW (sensor for PAR relying on WWE) enabled monitoring of PAR accumulation and depletion in live mammalian cells in response to different stimuli, namely hydrogen peroxide treatment, UV irradiation and hyperthermia.
Identifiants
pubmed: 32679873
pii: ijms21145004
doi: 10.3390/ijms21145004
pmc: PMC7404130
pii:
doi:
Substances chimiques
Bacterial Proteins
0
Fluorescent Dyes
0
Luminescent Proteins
0
Recombinant Fusion Proteins
0
yellow fluorescent protein, Bacteria
0
Poly Adenosine Diphosphate Ribose
26656-46-2
RNF146 protein, human
EC 2.3.2.27
Ubiquitin-Protein Ligases
EC 2.3.2.27
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : Russian Science Support Foundation
ID : 20-65-46018
Organisme : Russian Foundation for Basic Research
ID : 16-04-01870
Organisme : Ministry of Science and Higher Education of the Russian Federation
ID : RFMEFI62117X0018
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