Nuclear Actin Polymerization Rapidly Mediates Replication Fork Remodeling Upon Stress by Limiting PrimPol Activity

Overview

Journal Nat Commun

Specialty Biology

Date 2023 Nov 28

PMID 38016948

Authors

Maria Dilia Palumbieri

Chiara Merigliano

Daniel Gonzalez-Acosta

Danina Kuster

Jana Krietsch

Henriette Stoy

Thomas von Kanel

Svenja Ulferts

Bettina Welter

Joel Frey

Cyril Doerdelmann

Andrea Sanchi

Robert Grosse

Irene Chiolo

Massimo Lopes

Affiliations

Soon will be listed here.

Abstract

Cells rapidly respond to replication stress actively slowing fork progression and inducing fork reversal. How replication fork plasticity is achieved in the context of nuclear organization is currently unknown. Using nuclear actin probes in living and fixed cells, we visualized nuclear actin filaments in unperturbed S phase and observed their rapid extension in number and length upon genotoxic treatments, frequently taking contact with replication factories. Chemically or genetically impairing nuclear actin polymerization shortly before these treatments prevents active fork slowing and abolishes fork reversal. Defective fork remodeling is linked to deregulated chromatin loading of PrimPol, which promotes unrestrained and discontinuous DNA synthesis and limits the recruitment of RAD51 and SMARCAL1 to nascent DNA. Moreover, defective nuclear actin polymerization upon mild replication interference induces chromosomal instability in a PRIMPOL-dependent manner. Hence, by limiting PrimPol activity, nuclear F-actin orchestrates replication fork plasticity and is a key molecular determinant in the rapid cellular response to genotoxic treatments.

Citing Articles

ATR-hippo drives force signaling to nuclear F-actin and links mechanotransduction to neurological disorders.

Chatzifrangkeskou M, Stanly T, Koennig D, Campos-Soares L, Eyres M, Hasson A Sci Adv. 2025; 11(7):eadr5683.

PMID: 39951537 PMC: 11827640. DOI: 10.1126/sciadv.adr5683.

TORC2 inhibition triggers yeast chromosome fragmentation through misregulated Base Excision Repair of clustered oxidation events.

Shimada K, Tarashev C, Bregenhorn S, Gerhold C, van Loon B, Roth G Nat Commun. 2024; 15(1):9908.

PMID: 39548071 PMC: 11568337. DOI: 10.1038/s41467-024-54142-z.

Loss of cytoplasmic actin filaments raises nuclear actin levels to drive INO80C-dependent chromosome fragmentation.

Hurst V, Gerhold C, Tarashev C, Challa K, Seeber A, Yamazaki S Nat Commun. 2024; 15(1):9910.

PMID: 39548059 PMC: 11568269. DOI: 10.1038/s41467-024-54141-0.

Telomere maintenance and the DNA damage response: a paradoxical alliance.

Harman A, Bryan T Front Cell Dev Biol. 2024; 12:1472906.

PMID: 39483338 PMC: 11524846. DOI: 10.3389/fcell.2024.1472906.

Replication forks associated with long nuclear actin filaments in mild stress conditions display increased dynamics.

Merigliano C, Palumbieri M, Lopes M, Chiolo I MicroPubl Biol. 2024; 2024.

PMID: 39149411 PMC: 11325199. DOI: 10.17912/micropub.biology.001259.

References

Quinet A, Carvajal-Maldonado D, Lemacon D, Vindigni A . DNA Fiber Analysis: Mind the Gap!. Methods Enzymol. 2017; 591:55-82. DOI: 10.1016/bs.mie.2017.03.019. View

Jacobs K, Doerdelmann C, Krietsch J, Gonzalez-Acosta D, Mathis N, Kushinsky S . Stress-triggered hematopoietic stem cell proliferation relies on PrimPol-mediated repriming. Mol Cell. 2022; 82(21):4176-4188.e8. PMC: 10251193. DOI: 10.1016/j.molcel.2022.09.009. View

Nagai S, Dubrana K, Tsai-Pflugfelder M, Davidson M, Roberts T, Brown G . Functional targeting of DNA damage to a nuclear pore-associated SUMO-dependent ubiquitin ligase. Science. 2008; 322(5901):597-602. PMC: 3518492. DOI: 10.1126/science.1162790. View

Saxena S, Zou L . Hallmarks of DNA replication stress. Mol Cell. 2022; 82(12):2298-2314. PMC: 9219557. DOI: 10.1016/j.molcel.2022.05.004. View

Posern G, Sotiropoulos A, Treisman R . Mutant actins demonstrate a role for unpolymerized actin in control of transcription by serum response factor. Mol Biol Cell. 2002; 13(12):4167-78. PMC: 138624. DOI: 10.1091/mbc.02-05-0068. View

Wang Y, Hariharan A, Bastianello G, Toyama Y, Shivashankar G, Foiani M . DNA damage causes rapid accumulation of phosphoinositides for ATR signaling. Nat Commun. 2017; 8(1):2118. PMC: 5730617. DOI: 10.1038/s41467-017-01805-9. View

Klages-Mundt N, Kumar A, Zhang Y, Kapoor P, Shen X . The Nature of Actin-Family Proteins in Chromatin-Modifying Complexes. Front Genet. 2018; 9:398. PMC: 6167448. DOI: 10.3389/fgene.2018.00398. View

Hetrick B, Han M, Helgeson L, Nolen B . Small molecules CK-666 and CK-869 inhibit actin-related protein 2/3 complex by blocking an activating conformational change. Chem Biol. 2013; 20(5):701-12. PMC: 3684959. DOI: 10.1016/j.chembiol.2013.03.019. View

Stoy H, Luethi J, Roessler F, Riemann J, Kaech A, Lopes M . Direct R-Loop Visualization on Genomic DNA by Native Automated Electron Microscopy. Methods Mol Biol. 2022; 2528:1-20. DOI: 10.1007/978-1-0716-2477-7_1. View

10.

Oshidari R, Strecker J, Chung D, Abraham K, Chan J, Damaren C . Nuclear microtubule filaments mediate non-linear directional motion of chromatin and promote DNA repair. Nat Commun. 2018; 9(1):2567. PMC: 6028458. DOI: 10.1038/s41467-018-05009-7. View

11.

Plessner M, Melak M, Chinchilla P, Baarlink C, Grosse R . Nuclear F-actin formation and reorganization upon cell spreading. J Biol Chem. 2015; 290(18):11209-16. PMC: 4416828. DOI: 10.1074/jbc.M114.627166. View

12.

Mijic S, Zellweger R, Chappidi N, Berti M, Jacobs K, Mutreja K . Replication fork reversal triggers fork degradation in BRCA2-defective cells. Nat Commun. 2017; 8(1):859. PMC: 5643541. DOI: 10.1038/s41467-017-01164-5. View

13.

Maya-Mendoza A, Moudry P, Merchut-Maya J, Lee M, Strauss R, Bartek J . High speed of fork progression induces DNA replication stress and genomic instability. Nature. 2018; 559(7713):279-284. DOI: 10.1038/s41586-018-0261-5. View

14.

Hurst V, Shimada K, Gasser S . Nuclear Actin and Actin-Binding Proteins in DNA Repair. Trends Cell Biol. 2019; 29(6):462-476. DOI: 10.1016/j.tcb.2019.02.010. View

15.

Berti M, Ray Chaudhuri A, Thangavel S, Gomathinayagam S, Kenig S, Vujanovic M . Human RECQ1 promotes restart of replication forks reversed by DNA topoisomerase I inhibition. Nat Struct Mol Biol. 2013; 20(3):347-54. PMC: 3897332. DOI: 10.1038/nsmb.2501. View

16.

Baarlink C, Plessner M, Sherrard A, Morita K, Misu S, Virant D . A transient pool of nuclear F-actin at mitotic exit controls chromatin organization. Nat Cell Biol. 2017; 19(12):1389-1399. DOI: 10.1038/ncb3641. View

17.

Caridi C, Plessner M, Grosse R, Chiolo I . Nuclear actin filaments in DNA repair dynamics. Nat Cell Biol. 2019; 21(9):1068-1077. PMC: 6736642. DOI: 10.1038/s41556-019-0379-1. View

18.

Ray Chaudhuri A, Callen E, Ding X, Gogola E, Duarte A, Lee J . Replication fork stability confers chemoresistance in BRCA-deficient cells. Nature. 2016; 535(7612):382-7. PMC: 4959813. DOI: 10.1038/nature18325. View

19.

Wang Y, Sherrard A, Zhao B, Melak M, Trautwein J, Kleinschnitz E . GPCR-induced calcium transients trigger nuclear actin assembly for chromatin dynamics. Nat Commun. 2019; 10(1):5271. PMC: 6872576. DOI: 10.1038/s41467-019-13322-y. View

20.

Gaggioli V, Lo C, Reveron-Gomez N, Jasencakova Z, Domenech H, Nguyen H . Dynamic de novo heterochromatin assembly and disassembly at replication forks ensures fork stability. Nat Cell Biol. 2023; 25(7):1017-1032. PMC: 10344782. DOI: 10.1038/s41556-023-01167-z. View