Van Nerom, Margot and Ahmed, Junaid and Lázár, Tamás and Mészáros, Attila and Galand, Quentin and De Malsche, Wim and Van Lindt, Joris and Pancsa, Rita and Maes, Dominique and Tompa, Péter (2024) C9orf72-linked arginine-rich dipeptide repeats aggravate pathological phase separation of G3BP1. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 121 (50). No.-e2402847121. ISSN 0027-8424
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van-nerom-et-al-2024-c9orf72-linked-arginine-rich-dipeptide-repeats-aggravate-pathological-phase-separation-of-g3bp1.pdf - Published Version Available under License Creative Commons Attribution Non-commercial No Derivatives. Download (3MB) | Preview |
Abstract
The toxic effects of C9orf72-derived arginine-rich dipeptide repeats (R-DPRs) on cellular stress granules in amyotrophic lateral sclerosis (ALS) and frontotemporal dementia remain unclear at the molecular level. Stress granules are formed through the switch of Ras GTPase-activating protein-binding protein 1 (G3BP1) by RNA from a closed inactive state to an open activated state, driving the formation of the organelle by liquid–liquid phase separation (LLPS). We show that R-DPRs bind G3BP1 a thousand times stronger than RNA and initiate LLPS much more effectively. Their pathogenic effect is underscored by the slow transition of R-DPR–G3BP1 droplets to aggregated, ThS-positive states that can recruit ALS-linked proteins hnRNPA1, hnRNPA2, and TDP-43. Deletion constructs and molecular simulations show that R-DPR binding and LLPS are mediated via the negatively charged intrinsically disordered region 1 (IDR1) of the protein, allosterically regulated by its positively charged IDR3. Bioinformatic analyses point to the strong mechanistic parallels of these effects with the interaction of R-DPRs with nucleolar nucleophosmin 1 (NPM1) and underscore that R-DPRs interact with many other similar nucleolar and stress-granule proteins, extending the underlying mechanism of R-DPR toxicity in cells. Our results also highlight characteristic differences between the two R-DPRs, poly-GR and poly-PR, and suggest that the primary pathological target of poly-GR is not NPM1 in nucleoli, but G3BP1 in stress granules in affected cells.
| Item Type: | Article |
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| Additional Information: | Structural Biology Brussels, Bio-engineering Department, Vrije Universiteit Brussel, Elsene, 1050, Belgium Center for Structural Biology, Flanders Institute for Biotechnology, Elsene, 1050, Belgium Department of Chemical Engineering, Vrije Universiteit Brussel, Elsene, 1050, Belgium Institute of Molecular Life Sciences, Hungarian Research Network, Research Centre for Natural Sciences, Budapest, 1117, Hungary Export Date: 18 February 2025 CODEN: PNASA Correspondence Address: Tompa, P.; Structural Biology Brussels, Belgium; email: peter.tompa@vub.be Chemicals/CAS: arginine, 1119-34-2, 15595-35-4, 7004-12-8, 74-79-3; guanosine triphosphatase, 9059-32-9; nucleophosmin, 117896-08-9; DNA helicase; interferon induced helicase C domain containing protein 1; RNA helicase; transcriptional regulator ATRX; Werner syndrome ATP dependent helicase; Arginine; C9orf72 Protein; C9orf72 protein, human; Dipeptides; DNA Helicases; DNA-Binding Proteins; G3BP1 protein, human; Heterogeneous Nuclear Ribonucleoprotein A1; hnRNPA1 protein, human; NPM1 protein, human; Nucleophosmin; Poly-ADP-Ribose Binding Proteins; RNA Helicases; RNA Recognition Motif Proteins; TARDBP protein, human Manufacturers: Leica, Germany; Sartorius, Germany; Fluigent, France; Wyatt Technology Corp, United States Funding details: Agentschap Innoveren en Ondernemen, VLAIO Funding details: Tempus Közalapítvány, TPF Funding details: Fonds Wetenschappelijk Onderzoek, FWO, 11D2522N, FWOSB77 Funding details: Fonds Wetenschappelijk Onderzoek, FWO Funding details: Nemzeti Kutatási Fejlesztési és Innovációs Hivatal, NKFIH, BO/00174/22 Funding details: Nemzeti Kutatási Fejlesztési és Innovációs Hivatal, NKFIH Funding details: FK128133, K124670, K131702, FK142285, 778247 Funding details: European Space Agency, ESA, A0-2004-070 Funding details: European Space Agency, ESA Funding details: Magyar Tudományos Akadémia, MTA, 184018 Funding details: Magyar Tudományos Akadémia, MTA Funding details: Vrije Universiteit Brussel, VUB, SRP97, SRP51 Funding details: Vrije Universiteit Brussel, VUB Funding details: 952334 Funding text 1: We personally thank Prof. Paul Taylor (St. Jude Children's Research Hospital, Memphis, TN) for providing the construct of full-length G3BP1, and Antonio Maisto (Vrije Universiteit Brussel (VUB)) for familiarization with microfluidic experiments. We would like also to acknowledge funding from the following sources: EC H2020-WIDESPREAD-2020-5 Twinning grant (PhasAge, no. 952334, to P.T.), and EC H2020-MSCA-RISE Action grant (IDPfun, no. 778247, to P.T.); grants K124670 and K131702 (to P.T.) and FK128133 and FK142285 (to R.P.) from the National Research, Development and Innovation Office (NKFIH), Hungary; Bolyai fellowship BO/00174/22 (to R.P.) from the Hungarian Academy of Sciences; E\\u00F6tv\\u00F6s Research Fellowship no. 184018 (to R.P.) from the Tempus Public Foundation; VUB Strategic Research Programs (SRP51 and SRP97) at Vrije Universiteit Brussel (VUB, Brussels, to M.V.N., D.M., W.D.M., and P.T.); European Space Agency (ESA) grant A0-2004-070 (to D.M. and Q.G.); Fonds Wetenschappelijk Onderzoek (FWO) PhD fellowships in strategic basic research (FWOSB77, to J.A. and 11D2522N, to J.V.L.); postdoctoral innovation mandate (HBC.2022.0194) by the Flanders Innovation & Entrepreneurship Agency (VLAIO, to T.L.). |
| Uncontrolled Keywords: | G3BP1 , liquid–liquid phase separation , stress granules , neurodegenerative disease , repeat-expansion disease |
| Subjects: | Q Science / természettudomány > QH Natural history / természetrajz > QH301 Biology / biológia |
| SWORD Depositor: | MTMT SWORD |
| Depositing User: | MTMT SWORD |
| Date Deposited: | 16 Sep 2025 10:59 |
| Last Modified: | 16 Sep 2025 10:59 |
| URI: | https://real.mtak.hu/id/eprint/224315 |
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