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DNase I Induces Other Endonucleases in Kidney Tubular Epithelial Cells by Its DNA-Degrading Activity

Endonuclease-mediated DNA fragmentation is both an immediate cause and a result of apoptosis and of all other types of irreversible cell death after injury. It is produced by nine enzymes including DNase I, DNase 2, their homologs, caspase-activated DNase (CAD) and endonuclease G (EndoG). The endonu...

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Autores principales: Fahmi, Tariq, Wang, Xiaoying, Zhdanov, Dmitry D., Islam, Intisar, Apostolov, Eugene O., Savenka, Alena V., Basnakian, Alexei G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7698339/
https://www.ncbi.nlm.nih.gov/pubmed/33212932
http://dx.doi.org/10.3390/ijms21228665
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author Fahmi, Tariq
Wang, Xiaoying
Zhdanov, Dmitry D.
Islam, Intisar
Apostolov, Eugene O.
Savenka, Alena V.
Basnakian, Alexei G.
author_facet Fahmi, Tariq
Wang, Xiaoying
Zhdanov, Dmitry D.
Islam, Intisar
Apostolov, Eugene O.
Savenka, Alena V.
Basnakian, Alexei G.
author_sort Fahmi, Tariq
collection PubMed
description Endonuclease-mediated DNA fragmentation is both an immediate cause and a result of apoptosis and of all other types of irreversible cell death after injury. It is produced by nine enzymes including DNase I, DNase 2, their homologs, caspase-activated DNase (CAD) and endonuclease G (EndoG). The endonucleases act simultaneously during cell death; however, regulatory links between these enzymes have not been established. We hypothesized that DNase I, the most abundant of endonucleases, may regulate other endonucleases. To test this hypothesis, rat kidney tubular epithelial NRK-52E cells were transfected with the DNase I gene or its inactive mutant in a pECFP expression vector, while control cells were transfected with the empty vector. mRNA expression of all nine endonucleases was studied using real-time RT-PCR; DNA strand breaks in endonuclease genes were determined by PCR and protein expression of the enzymes was measured by Western blotting and quantitative immunocytochemistry. Our data showed that DNase I, but not its inactive mutant, induces all other endonucleases at varying time periods after transfection, causes DNA breaks in endonuclease genes, and elevates protein expression of several endonucleases. This is the first evidence that endonucleases seem to be induced by the DNA-degrading activity of DNase I.
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spelling pubmed-76983392020-11-29 DNase I Induces Other Endonucleases in Kidney Tubular Epithelial Cells by Its DNA-Degrading Activity Fahmi, Tariq Wang, Xiaoying Zhdanov, Dmitry D. Islam, Intisar Apostolov, Eugene O. Savenka, Alena V. Basnakian, Alexei G. Int J Mol Sci Article Endonuclease-mediated DNA fragmentation is both an immediate cause and a result of apoptosis and of all other types of irreversible cell death after injury. It is produced by nine enzymes including DNase I, DNase 2, their homologs, caspase-activated DNase (CAD) and endonuclease G (EndoG). The endonucleases act simultaneously during cell death; however, regulatory links between these enzymes have not been established. We hypothesized that DNase I, the most abundant of endonucleases, may regulate other endonucleases. To test this hypothesis, rat kidney tubular epithelial NRK-52E cells were transfected with the DNase I gene or its inactive mutant in a pECFP expression vector, while control cells were transfected with the empty vector. mRNA expression of all nine endonucleases was studied using real-time RT-PCR; DNA strand breaks in endonuclease genes were determined by PCR and protein expression of the enzymes was measured by Western blotting and quantitative immunocytochemistry. Our data showed that DNase I, but not its inactive mutant, induces all other endonucleases at varying time periods after transfection, causes DNA breaks in endonuclease genes, and elevates protein expression of several endonucleases. This is the first evidence that endonucleases seem to be induced by the DNA-degrading activity of DNase I. MDPI 2020-11-17 /pmc/articles/PMC7698339/ /pubmed/33212932 http://dx.doi.org/10.3390/ijms21228665 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
Fahmi, Tariq
Wang, Xiaoying
Zhdanov, Dmitry D.
Islam, Intisar
Apostolov, Eugene O.
Savenka, Alena V.
Basnakian, Alexei G.
DNase I Induces Other Endonucleases in Kidney Tubular Epithelial Cells by Its DNA-Degrading Activity
title DNase I Induces Other Endonucleases in Kidney Tubular Epithelial Cells by Its DNA-Degrading Activity
title_full DNase I Induces Other Endonucleases in Kidney Tubular Epithelial Cells by Its DNA-Degrading Activity
title_fullStr DNase I Induces Other Endonucleases in Kidney Tubular Epithelial Cells by Its DNA-Degrading Activity
title_full_unstemmed DNase I Induces Other Endonucleases in Kidney Tubular Epithelial Cells by Its DNA-Degrading Activity
title_short DNase I Induces Other Endonucleases in Kidney Tubular Epithelial Cells by Its DNA-Degrading Activity
title_sort dnase i induces other endonucleases in kidney tubular epithelial cells by its dna-degrading activity
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7698339/
https://www.ncbi.nlm.nih.gov/pubmed/33212932
http://dx.doi.org/10.3390/ijms21228665
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