Versatile enhancement of the killing potential of anti-cancer agents achieved by peptide mimetics of the PCNA interface towards specialized DNA polymerases

Abstract Cancer cells that survive chemotherapy achieve full DNA duplication despite the accumulation of damaged DNA triggered by chemotherapy. This happens because the synthesis of DNA at damaged sites is granted by tolerance events including translesion DNA synthesis (TLS), a process that promotes...

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Main Authors: Yiovana Verónica Okraine, María Belén de la Vega, Sofía Venerus Arbilla, Ginette Moyano, Agostina P. Bertolin, Horacio M. Pallarés, Lisa Wiesmüller, Sabrina F. Mansilla, Vanesa Gottifredi
Format: Article
Language:English
Published: Nature Publishing Group 2025-07-01
Series:Cell Death and Disease
Online Access:https://doi.org/10.1038/s41419-025-07812-9
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author Yiovana Verónica Okraine
María Belén de la Vega
Sofía Venerus Arbilla
Ginette Moyano
Agostina P. Bertolin
Horacio M. Pallarés
Lisa Wiesmüller
Sabrina F. Mansilla
Vanesa Gottifredi
author_facet Yiovana Verónica Okraine
María Belén de la Vega
Sofía Venerus Arbilla
Ginette Moyano
Agostina P. Bertolin
Horacio M. Pallarés
Lisa Wiesmüller
Sabrina F. Mansilla
Vanesa Gottifredi
author_sort Yiovana Verónica Okraine
collection DOAJ
description Abstract Cancer cells that survive chemotherapy achieve full DNA duplication despite the accumulation of damaged DNA triggered by chemotherapy. This happens because the synthesis of DNA at damaged sites is granted by tolerance events including translesion DNA synthesis (TLS), a process that promotes the use of specialized DNA polymerases (S-Pols) for DNA synthesis. Such a crucial role of S-Pols in the promotion of damaged DNA replication prompted analyses of the cell killing effects of individual S-Pols inhibitors. Because S-Pols can compensate for each other, a global inhibition of S-Pols needs to be designed and tested. Given that S-Pols are recruited to the replisome through their PCNA binding motif, we reasoned that global displacement of S-Pols will occur when delivering a peptide with a strong PCNA binding motif. The cyclin kinase inhibitor p21 contains the strongest PCNA binding motif. Therefore, we designed a peptide representing this C-terminal, PCNA interacting region (PIR) of p21. As hypothesized by us, the PIR peptide achieved global S-Pol displacement from PCNA-associated replication factories and enhanced the cancer cell killing potential of DNA damaging agents including cisplatin, hydroxyurea, olaparib and UV irradiation. Demonstrating strong versatility, the peptide also enhanced the cytotoxicity caused by agents that do not directly provoke DNA damage such as Chk1, ATR and Wee1 inhibitors. In all cases, disrupting the PCNA binding site within the PIR peptide was sufficient to dismantle its cell killing potential. Strengthening the concept, a less potent PIR, namely derived from S-Pol eta, efficiently displaced S-Pols from replication factories exacerbating cell killing by all agents tested. These results collectively indicate that simultaneous displacement of S-Pols from PCNA can be enforced by excess levels of PIR peptides. This strategy is demonstrably valid to enhance the cancer cell killing by different DNA-damaging agents.
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spelling doaj-art-b4dd400ec95d462aa0b44d4c199369282025-08-20T03:06:09ZengNature Publishing GroupCell Death and Disease2041-48892025-07-0116111710.1038/s41419-025-07812-9Versatile enhancement of the killing potential of anti-cancer agents achieved by peptide mimetics of the PCNA interface towards specialized DNA polymerasesYiovana Verónica Okraine0María Belén de la Vega1Sofía Venerus Arbilla2Ginette Moyano3Agostina P. Bertolin4Horacio M. Pallarés5Lisa Wiesmüller6Sabrina F. Mansilla7Vanesa Gottifredi8Fundación Instituto Leloir—Instituto de Investigaciones Bioquímicas de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)Fundación Instituto Leloir—Instituto de Investigaciones Bioquímicas de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)Fundación Instituto Leloir—Instituto de Investigaciones Bioquímicas de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)Fundación Instituto Leloir—Instituto de Investigaciones Bioquímicas de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)Fundación Instituto Leloir—Instituto de Investigaciones Bioquímicas de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)Fundación Instituto Leloir—Instituto de Investigaciones Bioquímicas de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)Department of Obstetrics and Gynecology, Ulm UniversityFundación Instituto Leloir—Instituto de Investigaciones Bioquímicas de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)Fundación Instituto Leloir—Instituto de Investigaciones Bioquímicas de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)Abstract Cancer cells that survive chemotherapy achieve full DNA duplication despite the accumulation of damaged DNA triggered by chemotherapy. This happens because the synthesis of DNA at damaged sites is granted by tolerance events including translesion DNA synthesis (TLS), a process that promotes the use of specialized DNA polymerases (S-Pols) for DNA synthesis. Such a crucial role of S-Pols in the promotion of damaged DNA replication prompted analyses of the cell killing effects of individual S-Pols inhibitors. Because S-Pols can compensate for each other, a global inhibition of S-Pols needs to be designed and tested. Given that S-Pols are recruited to the replisome through their PCNA binding motif, we reasoned that global displacement of S-Pols will occur when delivering a peptide with a strong PCNA binding motif. The cyclin kinase inhibitor p21 contains the strongest PCNA binding motif. Therefore, we designed a peptide representing this C-terminal, PCNA interacting region (PIR) of p21. As hypothesized by us, the PIR peptide achieved global S-Pol displacement from PCNA-associated replication factories and enhanced the cancer cell killing potential of DNA damaging agents including cisplatin, hydroxyurea, olaparib and UV irradiation. Demonstrating strong versatility, the peptide also enhanced the cytotoxicity caused by agents that do not directly provoke DNA damage such as Chk1, ATR and Wee1 inhibitors. In all cases, disrupting the PCNA binding site within the PIR peptide was sufficient to dismantle its cell killing potential. Strengthening the concept, a less potent PIR, namely derived from S-Pol eta, efficiently displaced S-Pols from replication factories exacerbating cell killing by all agents tested. These results collectively indicate that simultaneous displacement of S-Pols from PCNA can be enforced by excess levels of PIR peptides. This strategy is demonstrably valid to enhance the cancer cell killing by different DNA-damaging agents.https://doi.org/10.1038/s41419-025-07812-9
spellingShingle Yiovana Verónica Okraine
María Belén de la Vega
Sofía Venerus Arbilla
Ginette Moyano
Agostina P. Bertolin
Horacio M. Pallarés
Lisa Wiesmüller
Sabrina F. Mansilla
Vanesa Gottifredi
Versatile enhancement of the killing potential of anti-cancer agents achieved by peptide mimetics of the PCNA interface towards specialized DNA polymerases
Cell Death and Disease
title Versatile enhancement of the killing potential of anti-cancer agents achieved by peptide mimetics of the PCNA interface towards specialized DNA polymerases
title_full Versatile enhancement of the killing potential of anti-cancer agents achieved by peptide mimetics of the PCNA interface towards specialized DNA polymerases
title_fullStr Versatile enhancement of the killing potential of anti-cancer agents achieved by peptide mimetics of the PCNA interface towards specialized DNA polymerases
title_full_unstemmed Versatile enhancement of the killing potential of anti-cancer agents achieved by peptide mimetics of the PCNA interface towards specialized DNA polymerases
title_short Versatile enhancement of the killing potential of anti-cancer agents achieved by peptide mimetics of the PCNA interface towards specialized DNA polymerases
title_sort versatile enhancement of the killing potential of anti cancer agents achieved by peptide mimetics of the pcna interface towards specialized dna polymerases
url https://doi.org/10.1038/s41419-025-07812-9
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