Genetically Encoded Fluorescent Sensor for Poly-ADP-Ribose

被引：6

作者：

Serebrovskaya, Ekaterina O. ^{[1
]}

Podvalnaya, Nadezda M. ^{[1
,2
]}

Dudenkova, Varvara V. ^{[3
]}

Efremova, Anna S. ^{[2
]}

Gurskaya, Nadya G. ^{[1
,4
]}

Gorbachev, Dmitry A. ^{[1
]}

Luzhin, Artem, V ^{[5
]}

Kantidze, Omar L. ^{[5
]}

Zagaynova, Elena, V ^{[3
,6
]}

Shram, Stanislav, I ^{[2
]}

Lukyanov, Konstantin A. ^{[1
]}

机构：

[1] Shemyakin Ovchinnikov Inst Bioorgan Chem, Miklukho Maklaya 16-10, Moscow 117997, Russia

[2] Inst Mol Genet, Kurchatova Sq 2, Moscow 123182, Russia

[3] Privolzhsky Res Med Univ, Inst Expt Oncol & Biomed Technol, Minin & Pozharsky Sq 10-1, Nizhnii Novgorod 603005, Russia

[4] Pirogov Russian Natl Res Med Univ, Inst Translat Med, Ostrovityanova 1, Moscow 117997, Russia

[5] Inst Gene Biol, Vavilova 34-5, Moscow 119334, Russia

[6] Lobachevsky State Univ Nizhny Novgorod, Gagarin Ave 23, Nizhnii Novgorod 603950, Russia

来源：

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES | 2020年 / 21卷 / 14期

基金：

俄罗斯科学基金会; 俄罗斯基础研究基金会;

关键词：

DNA damage respose; WWE-domai; PAR; fluorescent protein; FRET; sensor; POLY(ADP-RIBOSE); MECHANISM; INSIGHTS; PROTEIN;

D O I：

10.3390/ijms21145004

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 Forster 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 forPARrelyingonWWE) enabled monitoring of PAR accumulation and depletion in live mammalian cells in response to different stimuli, namely hydrogen peroxide treatment, UV irradiation and hyperthermia.

引用

页码：1 / 11

页数：11

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