TY - JOUR

T1 - PARP-1 Activation Directs FUS to DNA Damage Sites to Form PARG-Reversible Compartments Enriched in Damaged DNA

AU - Singatulina, Anastasia S.

AU - Hamon, Loic

AU - Sukhanova, Maria V.

AU - Desforges, Bénédicte

AU - Joshi, Vandana

AU - Bouhss, Ahmed

AU - Lavrik, Olga I.

AU - Pastré, David

N1 - Publisher Copyright: © 2019 The Authors

PY - 2019/5/7

Y1 - 2019/5/7

N2 - PARP-1 synthesizes long poly(ADP-ribose) chains (PAR) at DNA damage sites to recruit DNA repair factors. Among proteins relocated on damaged DNA, the RNA-binding protein FUS is one of the most abundant, raising the issue about its involvement in DNA repair. Here, we reconstituted the PARP-1/PAR/DNA system in vitro and analyzed at the single-molecule level the role of FUS. We demonstrate successively the dissociation of FUS from mRNA, its recruitment at DNA damage sites through its binding to PAR, and the assembly of damaged DNA-rich compartments. PARG, an enzyme family that hydrolyzes PAR, is sufficient to dissociate damaged DNA-rich compartments in vitro and initiates the nucleocytoplasmic shuttling of FUS in cells. We anticipate that, consistent with previous models, FUS facilitates DNA repair through the transient compartmentalization of DNA damage sites. The nucleocytoplasmic shuttling of FUS after the PARG-mediated compartment dissociation may participate in the formation of cytoplasmic FUS aggregates. Using a single-molecule approach, Singatulina et al. reconstitute the DNA repair system leading to the specific recruitment of FUS, an mRNA-binding protein related to liquid-liquid phase separation biology, to DNA damage sites, thus revealing the capacity of FUS to form dynamic compartments in which damaged DNA is concentrated.

AB - PARP-1 synthesizes long poly(ADP-ribose) chains (PAR) at DNA damage sites to recruit DNA repair factors. Among proteins relocated on damaged DNA, the RNA-binding protein FUS is one of the most abundant, raising the issue about its involvement in DNA repair. Here, we reconstituted the PARP-1/PAR/DNA system in vitro and analyzed at the single-molecule level the role of FUS. We demonstrate successively the dissociation of FUS from mRNA, its recruitment at DNA damage sites through its binding to PAR, and the assembly of damaged DNA-rich compartments. PARG, an enzyme family that hydrolyzes PAR, is sufficient to dissociate damaged DNA-rich compartments in vitro and initiates the nucleocytoplasmic shuttling of FUS in cells. We anticipate that, consistent with previous models, FUS facilitates DNA repair through the transient compartmentalization of DNA damage sites. The nucleocytoplasmic shuttling of FUS after the PARG-mediated compartment dissociation may participate in the formation of cytoplasmic FUS aggregates. Using a single-molecule approach, Singatulina et al. reconstitute the DNA repair system leading to the specific recruitment of FUS, an mRNA-binding protein related to liquid-liquid phase separation biology, to DNA damage sites, thus revealing the capacity of FUS to form dynamic compartments in which damaged DNA is concentrated.

KW - atomic force microscopy

KW - cancer

KW - DNA repair

KW - liquid-liquid phase separation

KW - neurodegenerative disease

KW - poly(ADP-ribose)

KW - poly(ADP-ribose) polymerase 1

KW - RNA-binding proteins

KW - NUCLEAR IMPORT

KW - POLY(ADP-RIBOSE)

KW - DEPENDENT RECRUITMENT

KW - ALS

KW - PHASE-SEPARATION

KW - MESSENGER-RNA

KW - ARGININE METHYLATION

KW - STRESS

KW - PROTEINS

KW - BINDING

UR - http://www.scopus.com/inward/record.url?scp=85064597303&partnerID=8YFLogxK

U2 - 10.1016/j.celrep.2019.04.031

DO - 10.1016/j.celrep.2019.04.031

M3 - Article

C2 - 31067465

AN - SCOPUS:85064597303

VL - 27

SP - 1809-1821.e5

JO - Cell Reports

JF - Cell Reports

SN - 2211-1247

IS - 6

ER -