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Yeast Rpn4 Links the Proteasome and DNA Repair via RAD52 Regulation

Environmental and intracellular factors often damage DNA, but multiple DNA repair pathways maintain genome integrity. In yeast, the 26S proteasome and its transcriptional regulator and substrate Rpn4 are involved in DNA damage resistance. Paradoxically, while proteasome dysfunction may induce hyper-...

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Autores principales: Spasskaya, Daria S., Nadolinskaia, Nonna I., Tutyaeva, Vera V., Lysov, Yuriy P., Karpov, Vadim L., Karpov, Dmitry S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7672625/
https://www.ncbi.nlm.nih.gov/pubmed/33143019
http://dx.doi.org/10.3390/ijms21218097
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author Spasskaya, Daria S.
Nadolinskaia, Nonna I.
Tutyaeva, Vera V.
Lysov, Yuriy P.
Karpov, Vadim L.
Karpov, Dmitry S.
author_facet Spasskaya, Daria S.
Nadolinskaia, Nonna I.
Tutyaeva, Vera V.
Lysov, Yuriy P.
Karpov, Vadim L.
Karpov, Dmitry S.
author_sort Spasskaya, Daria S.
collection PubMed
description Environmental and intracellular factors often damage DNA, but multiple DNA repair pathways maintain genome integrity. In yeast, the 26S proteasome and its transcriptional regulator and substrate Rpn4 are involved in DNA damage resistance. Paradoxically, while proteasome dysfunction may induce hyper-resistance to DNA-damaging agents, Rpn4 malfunction sensitizes yeasts to these agents. Previously, we proposed that proteasome inhibition causes Rpn4 stabilization followed by the upregulation of Rpn4-dependent DNA repair genes and pathways. Here, we aimed to elucidate the key Rpn4 targets responsible for DNA damage hyper-resistance in proteasome mutants. We impaired the Rpn4-mediated regulation of candidate genes using the CRISPR/Cas9 system and tested the sensitivity of mutant strains to 4-NQO, MMS and zeocin. We found that the separate or simultaneous deregulation of 19S or 20S proteasome subcomplexes induced MAG1, DDI1, RAD23 and RAD52 in an Rpn4-dependent manner. Deregulation of RAD23, DDI1 and RAD52 sensitized yeast to DNA damage. Genetic, epigenetic or dihydrocoumarin-mediated RAD52 repression restored the sensitivity of the proteasome mutants to DNA damage. Our results suggest that the Rpn4-mediated overexpression of DNA repair genes, especially RAD52, defines the DNA damage hyper-resistant phenotype of proteasome mutants. The developed yeast model is useful for characterizing drugs that reverse the DNA damage hyper-resistance phenotypes of cancers.
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spelling pubmed-76726252020-11-19 Yeast Rpn4 Links the Proteasome and DNA Repair via RAD52 Regulation Spasskaya, Daria S. Nadolinskaia, Nonna I. Tutyaeva, Vera V. Lysov, Yuriy P. Karpov, Vadim L. Karpov, Dmitry S. Int J Mol Sci Article Environmental and intracellular factors often damage DNA, but multiple DNA repair pathways maintain genome integrity. In yeast, the 26S proteasome and its transcriptional regulator and substrate Rpn4 are involved in DNA damage resistance. Paradoxically, while proteasome dysfunction may induce hyper-resistance to DNA-damaging agents, Rpn4 malfunction sensitizes yeasts to these agents. Previously, we proposed that proteasome inhibition causes Rpn4 stabilization followed by the upregulation of Rpn4-dependent DNA repair genes and pathways. Here, we aimed to elucidate the key Rpn4 targets responsible for DNA damage hyper-resistance in proteasome mutants. We impaired the Rpn4-mediated regulation of candidate genes using the CRISPR/Cas9 system and tested the sensitivity of mutant strains to 4-NQO, MMS and zeocin. We found that the separate or simultaneous deregulation of 19S or 20S proteasome subcomplexes induced MAG1, DDI1, RAD23 and RAD52 in an Rpn4-dependent manner. Deregulation of RAD23, DDI1 and RAD52 sensitized yeast to DNA damage. Genetic, epigenetic or dihydrocoumarin-mediated RAD52 repression restored the sensitivity of the proteasome mutants to DNA damage. Our results suggest that the Rpn4-mediated overexpression of DNA repair genes, especially RAD52, defines the DNA damage hyper-resistant phenotype of proteasome mutants. The developed yeast model is useful for characterizing drugs that reverse the DNA damage hyper-resistance phenotypes of cancers. MDPI 2020-10-30 /pmc/articles/PMC7672625/ /pubmed/33143019 http://dx.doi.org/10.3390/ijms21218097 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Spasskaya, Daria S.
Nadolinskaia, Nonna I.
Tutyaeva, Vera V.
Lysov, Yuriy P.
Karpov, Vadim L.
Karpov, Dmitry S.
Yeast Rpn4 Links the Proteasome and DNA Repair via RAD52 Regulation
title Yeast Rpn4 Links the Proteasome and DNA Repair via RAD52 Regulation
title_full Yeast Rpn4 Links the Proteasome and DNA Repair via RAD52 Regulation
title_fullStr Yeast Rpn4 Links the Proteasome and DNA Repair via RAD52 Regulation
title_full_unstemmed Yeast Rpn4 Links the Proteasome and DNA Repair via RAD52 Regulation
title_short Yeast Rpn4 Links the Proteasome and DNA Repair via RAD52 Regulation
title_sort yeast rpn4 links the proteasome and dna repair via rad52 regulation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7672625/
https://www.ncbi.nlm.nih.gov/pubmed/33143019
http://dx.doi.org/10.3390/ijms21218097
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