{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,20]],"date-time":"2026-04-20T19:58:19Z","timestamp":1776715099243,"version":"3.51.2"},"reference-count":49,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2020,2,18]],"date-time":"2020-02-18T00:00:00Z","timestamp":1581984000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001824","name":"Grantov\u00e1 Agentura \u010cesk\u00e9 Republiky","doi-asserted-by":"publisher","award":["GACR 17-25976S"],"award-info":[{"award-number":["GACR 17-25976S"]}],"id":[{"id":"10.13039\/501100001824","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Ministry of School, Education, Youth and Sports of the Czech Republic","award":["LM2015062 Czech\u2010BioImaging"],"award-info":[{"award-number":["LM2015062 Czech\u2010BioImaging"]}]},{"name":"Ministry of School, Education, Youth and Sports of the Czech Republic","award":["ENOCH No. CZ.02.1.01\/0.0\/0.0\/16_019\/0000868"],"award-info":[{"award-number":["ENOCH No. CZ.02.1.01\/0.0\/0.0\/16_019\/0000868"]}]},{"name":"Internal grant of the Palacky University","award":["IGA_LF_2019_026"],"award-info":[{"award-number":["IGA_LF_2019_026"]}]},{"name":"the Novo Nordisk Foundation","award":["16854"],"award-info":[{"award-number":["16854"]}]},{"name":"the Danish National Research Foundation","award":["project CARD: no. DNRF125"],"award-info":[{"award-number":["project CARD: no. DNRF125"]}]},{"DOI":"10.13039\/100008363","name":"the Danish Cancer Society","doi-asserted-by":"publisher","award":["R204-A12617"],"award-info":[{"award-number":["R204-A12617"]}],"id":[{"id":"10.13039\/100008363","id-type":"DOI","asserted-by":"publisher"}]},{"name":"the Swedish Research Council","award":["VR-MH 2014-46602-117891-30"],"award-info":[{"award-number":["VR-MH 2014-46602-117891-30"]}]},{"DOI":"10.13039\/501100002794","name":"Swedish Cancer Society","doi-asserted-by":"publisher","award":["170176"],"award-info":[{"award-number":["170176"]}],"id":[{"id":"10.13039\/501100002794","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>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.<\/jats:p>","DOI":"10.3390\/cells9020469","type":"journal-article","created":{"date-parts":[[2020,2,25]],"date-time":"2020-02-25T04:21:26Z","timestamp":1582604486000},"page":"469","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":43,"title":["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"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9238-3437","authenticated-orcid":false,"given":"Dusana","family":"Majera","sequence":"first","affiliation":[{"name":"Laboratory of Genome Integrity, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, 77 147 Olomouc, Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6004-3609","authenticated-orcid":false,"given":"Zdenek","family":"Skrott","sequence":"additional","affiliation":[{"name":"Laboratory of Genome Integrity, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, 77 147 Olomouc, Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0141-0469","authenticated-orcid":false,"given":"Katarina","family":"Chroma","sequence":"additional","affiliation":[{"name":"Laboratory of Genome Integrity, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, 77 147 Olomouc, Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joanna Maria","family":"Merchut-Maya","sequence":"additional","affiliation":[{"name":"Danish Cancer Society Research Center, 2100 Copenhagen, Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2321-0348","authenticated-orcid":false,"given":"Martin","family":"Mistrik","sequence":"additional","affiliation":[{"name":"Laboratory of Genome Integrity, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, 77 147 Olomouc, Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiri","family":"Bartek","sequence":"additional","affiliation":[{"name":"Laboratory of Genome Integrity, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, 77 147 Olomouc, Czech Republic"},{"name":"Danish Cancer Society Research Center, 2100 Copenhagen, Denmark"},{"name":"Division of Genome Biology, Department of Medical Biochemistry and Biophysics, Science for Life Laboratory, Karolinska Institute, 171 77 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,2,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1038\/nrd3461","article-title":"Mining for therapeutic gold","volume":"10","author":"Collins","year":"2011","journal-title":"Nat. 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