Targeting the NPL4 Adaptor of P97/VCP Segregase by Disulfiram As an Emerging Cancer Vulnerability Evokes Replication Stress and DNA Damage While Silencing the ATR Pathway

Overview

Journal Cells

Publisher MDPI

Specialties Biochemistry
Biophysics
Cell Biology
Molecular Biology

Date 2020 Feb 23

PMID 32085572

Citations 23

Authors

Dusana Majera

Zdenek Skrott

Katarina Chroma

Joanna Maria Merchut-Maya

Martin Mistrik

Jiri Bartek

Affiliations

Soon will be listed here.

Abstract

Research on repurposing the old alcohol-aversion drug disulfiram (DSF) for cancer treatment has identified inhibition of NPL4, an adaptor of the p97/VCP segregase essential for turnover of proteins involved in multiple pathways, as an unsuspected cancer cell vulnerability. While we reported that NPL4 is targeted by the anticancer metabolite of DSF, the bis-diethyldithiocarbamate-copper complex (CuET), the exact, apparently multifaceted mechanism(s) through which the CuET-induced aggregation of NPL4 kills cancer cells remains to be fully elucidated. Given the pronounced sensitivity to CuET in tumor cell lines lacking the genome integrity caretaker proteins BRCA1 and BRCA2, here we investigated the impact of NPL4 targeting by CuET on DNA replication dynamics and DNA damage response pathways in human cancer cell models. Our results show that CuET treatment interferes with DNA replication, slows down replication fork progression and causes accumulation of single-stranded DNA (ssDNA). Such a replication stress (RS) scenario is associated with DNA damage, preferentially in the S phase, and activates the homologous recombination (HR) DNA repair pathway. At the same time, we find that cellular responses to the CuET-triggered RS are seriously impaired due to concomitant malfunction of the ATRIP-ATR-CHK1 signaling pathway that reflects an unorthodox checkpoint silencing mode through ATR (Ataxia telangiectasia and Rad3 related) kinase sequestration within the CuET-evoked NPL4 protein aggregates.

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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

Wang Y, Li W, Patel S, Cong J, Zhang N, Sabbatino F . Blocking the formation of radiation-induced breast cancer stem cells. Oncotarget. 2014; 5(11):3743-55. PMC: 4116517. DOI: 10.18632/oncotarget.1992. View

Davis E, Lachaud C, Appleton P, Macartney T, Nathke I, Rouse J . DVC1 (C1orf124) recruits the p97 protein segregase to sites of DNA damage. Nat Struct Mol Biol. 2012; 19(11):1093-100. DOI: 10.1038/nsmb.2394. View

Skrott Z, Majera D, Gursky J, Buchtova T, Hajduch M, Mistrik M . Disulfiram's anti-cancer activity reflects targeting NPL4, not inhibition of aldehyde dehydrogenase. Oncogene. 2019; 38(40):6711-6722. DOI: 10.1038/s41388-019-0915-2. View

Toledo L, Altmeyer M, Rask M, Lukas C, Larsen D, Povlsen L . ATR prohibits replication catastrophe by preventing global exhaustion of RPA. Cell. 2013; 155(5):1088-103. DOI: 10.1016/j.cell.2013.10.043. View

Bartkova J, Horejsi Z, Koed K, Kramer A, Tort F, Zieger K . DNA damage response as a candidate anti-cancer barrier in early human tumorigenesis. Nature. 2005; 434(7035):864-70. DOI: 10.1038/nature03482. View

Budzowska M, Kanaar R . Mechanisms of dealing with DNA damage-induced replication problems. Cell Biochem Biophys. 2008; 53(1):17-31. DOI: 10.1007/s12013-008-9039-y. View

Ramadan K, Halder S, Wiseman K, Vaz B . Strategic role of the ubiquitin-dependent segregase p97 (VCP or Cdc48) in DNA replication. Chromosoma. 2016; 126(1):17-32. DOI: 10.1007/s00412-016-0587-4. View

Bekker-Jensen S, Lukas C, Kitagawa R, Melander F, Kastan M, Bartek J . Spatial organization of the mammalian genome surveillance machinery in response to DNA strand breaks. J Cell Biol. 2006; 173(2):195-206. PMC: 2063811. DOI: 10.1083/jcb.200510130. View

10.

Singh A, Oehler J, Torrecilla I, Kilgas S, Li S, Vaz B . The p97-Ataxin 3 complex regulates homeostasis of the DNA damage response E3 ubiquitin ligase RNF8. EMBO J. 2019; 38(21):e102361. PMC: 6826192. DOI: 10.15252/embj.2019102361. View

11.

Lorenti Garcia C, Mechilli M, Proietti De Santis L, Schinoppi A, Kobos K, Palitti F . Relationship between DNA lesions, DNA repair and chromosomal damage induced by acetaldehyde. Mutat Res. 2008; 662(1-2):3-9. DOI: 10.1016/j.mrfmmm.2008.11.008. View

12.

Yasuda S, Tsuchiya H, Kaiho A, Guo Q, Ikeuchi K, Endo A . Stress- and ubiquitylation-dependent phase separation of the proteasome. Nature. 2020; 578(7794):296-300. DOI: 10.1038/s41586-020-1982-9. View

13.

Bergink S, Ammon T, Kern M, Schermelleh L, Leonhardt H, Jentsch S . Role of Cdc48/p97 as a SUMO-targeted segregase curbing Rad51-Rad52 interaction. Nat Cell Biol. 2013; 15(5):526-32. DOI: 10.1038/ncb2729. View

14.

Gorgoulis V, Vassiliou L, Karakaidos P, Zacharatos P, Kotsinas A, Liloglou T . Activation of the DNA damage checkpoint and genomic instability in human precancerous lesions. Nature. 2005; 434(7035):907-13. DOI: 10.1038/nature03485. View

15.

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

16.

Davies A, Masson J, McIlwraith M, Stasiak A, Stasiak A, Venkitaraman A . Role of BRCA2 in control of the RAD51 recombination and DNA repair protein. Mol Cell. 2001; 7(2):273-82. DOI: 10.1016/s1097-2765(01)00175-7. View

17.

Collins F . Mining for therapeutic gold. Nat Rev Drug Discov. 2011; 10(6):397. PMC: 5101935. DOI: 10.1038/nrd3461. View

18.

Tacconi E, Lai X, Folio C, Porru M, Zonderland G, Badie S . BRCA1 and BRCA2 tumor suppressors protect against endogenous acetaldehyde toxicity. EMBO Mol Med. 2017; 9(10):1398-1414. PMC: 5623864. DOI: 10.15252/emmm.201607446. View

19.

Liao H, Ji F, Helleday T, Ying S . Mechanisms for stalled replication fork stabilization: new targets for synthetic lethality strategies in cancer treatments. EMBO Rep. 2018; 19(9). PMC: 6123652. DOI: 10.15252/embr.201846263. View

20.

Jakola A, Werlenius K, Mudaisi M, Hylin S, Kinhult S, Bartek Jr J . Disulfiram repurposing combined with nutritional copper supplement as add-on to chemotherapy in recurrent glioblastoma (DIRECT): Study protocol for a randomized controlled trial. F1000Res. 2019; 7:1797. PMC: 6325620. DOI: 10.12688/f1000research.16786.1. View