CRL4 ubiquitin ligase stimulates Fanconi anemia pathway-induced single-stranded DNA-RPA signaling.
Antineoplastic Agents
/ pharmacology
Ataxia Telangiectasia Mutated Proteins
/ metabolism
Cell Cycle
Cell Line, Tumor
Cell Survival
Cisplatin
/ pharmacology
Cullin Proteins
/ genetics
DNA Damage
DNA, Single-Stranded
/ genetics
Fanconi Anemia
/ metabolism
Female
Humans
Mitomycin
/ pharmacology
Ovarian Neoplasms
/ drug therapy
RNA, Small Interfering
/ genetics
S Phase Cell Cycle Checkpoints
Signal Transduction
Ubiquitin-Protein Ligases
/ genetics
Ubiquitination
Chemotherapy
Cisplatin
Crosslink - CUL4 - Fanconi anemia
ssDNA
Journal
BMC cancer
ISSN: 1471-2407
Titre abrégé: BMC Cancer
Pays: England
ID NLM: 100967800
Informations de publication
Date de publication:
05 Nov 2019
05 Nov 2019
Historique:
received:
13
08
2019
accepted:
28
10
2019
entrez:
7
11
2019
pubmed:
7
11
2019
medline:
4
4
2020
Statut:
epublish
Résumé
DNA-crosslinking agents like cisplatin and mitomycin C (MMC) are indispensible for the treatment of many solid malignancies. These anticancer drugs generate DNA interstrand crosslinks (ICLs) that cause cell death by blocking replication forks. Many factors counteracting ICL-induced DNA replication stress, including the Fanconi anemia (FA) pathway, are regulated by ubiquitination and, therefore, ubiquitin ligases are potential targets for the sensitization of cancer cells to crosslinking agents. In this study, we investigated the function of the CRL4 ubiquitin ligase in modulating the response of cancer cells to ICL induction. The two cullin paralogs CUL4A and CUL4B, which form the CRL4 ligase scaffold, were depleted in cancer cells by small interfering RNA followed by analysis of the cellular and biochemical responses to ICLs elicited upon cisplatin or MMC treatment. We report that the combined depletion of CUL4A and CUL4B weakens an FA pathway-dependent S phase checkpoint response. CRL4 positively stimulates the monoubiquitination of FANCD2 required for the recruitment of XPF-ERCC1, a structure-specific endonuclease that, in turn, contributes to the display of single-stranded DNA (ssDNA) at ICLs. After CRL4 down regulation, the missing ssDNA results in reduced recruitment of RPA, thereby dampening activation of ATR and CHK1 checkpoint kinases and allowing for S phase progression despite ICL induction. Our findings indicate that CRL4 promotes cell survival by potentiating an FA pathway-dependent ssDNA-RPA signaling platform installed at ICLs. The anticancer efficacy of crosslinking agents may, therefore, be enhanced by down regulating CRL4 activity.
Sections du résumé
BACKGROUND
BACKGROUND
DNA-crosslinking agents like cisplatin and mitomycin C (MMC) are indispensible for the treatment of many solid malignancies. These anticancer drugs generate DNA interstrand crosslinks (ICLs) that cause cell death by blocking replication forks. Many factors counteracting ICL-induced DNA replication stress, including the Fanconi anemia (FA) pathway, are regulated by ubiquitination and, therefore, ubiquitin ligases are potential targets for the sensitization of cancer cells to crosslinking agents. In this study, we investigated the function of the CRL4 ubiquitin ligase in modulating the response of cancer cells to ICL induction.
METHODS
METHODS
The two cullin paralogs CUL4A and CUL4B, which form the CRL4 ligase scaffold, were depleted in cancer cells by small interfering RNA followed by analysis of the cellular and biochemical responses to ICLs elicited upon cisplatin or MMC treatment.
RESULTS
RESULTS
We report that the combined depletion of CUL4A and CUL4B weakens an FA pathway-dependent S phase checkpoint response. CRL4 positively stimulates the monoubiquitination of FANCD2 required for the recruitment of XPF-ERCC1, a structure-specific endonuclease that, in turn, contributes to the display of single-stranded DNA (ssDNA) at ICLs. After CRL4 down regulation, the missing ssDNA results in reduced recruitment of RPA, thereby dampening activation of ATR and CHK1 checkpoint kinases and allowing for S phase progression despite ICL induction.
CONCLUSION
CONCLUSIONS
Our findings indicate that CRL4 promotes cell survival by potentiating an FA pathway-dependent ssDNA-RPA signaling platform installed at ICLs. The anticancer efficacy of crosslinking agents may, therefore, be enhanced by down regulating CRL4 activity.
Identifiants
pubmed: 31690264
doi: 10.1186/s12885-019-6305-x
pii: 10.1186/s12885-019-6305-x
pmc: PMC6833152
doi:
Substances chimiques
Antineoplastic Agents
0
CUL4A protein, human
0
CUL4B protein, human
0
Cullin Proteins
0
DNA, Single-Stranded
0
IL17RB protein, human
0
RNA, Small Interfering
0
Mitomycin
50SG953SK6
Ubiquitin-Protein Ligases
EC 2.3.2.27
ATR protein, human
EC 2.7.11.1
Ataxia Telangiectasia Mutated Proteins
EC 2.7.11.1
Cisplatin
Q20Q21Q62J
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
1042Subventions
Organisme : Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung
ID : 170111/1
Organisme : Velux Stiftung
ID : 753
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