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p53 CRISPR Deletion Affects DNA Structure and Nuclear Architecture
The TP53 gene is a key tumor suppressor. Although the tumor suppressor p53 was one of the first to be characterized as a transcription factor, with its main function potentiated by its interaction with DNA, there are still many unresolved questions about its mechanism of action. Here, we demonstrate...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7073688/ https://www.ncbi.nlm.nih.gov/pubmed/32098416 http://dx.doi.org/10.3390/jcm9020598 |
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author | Rangel-Pozzo, Aline Booth, Samuel Yu, Pak Lok Ivan Singh, Madhurendra Selivanova, Galina Mai, Sabine |
author_facet | Rangel-Pozzo, Aline Booth, Samuel Yu, Pak Lok Ivan Singh, Madhurendra Selivanova, Galina Mai, Sabine |
author_sort | Rangel-Pozzo, Aline |
collection | PubMed |
description | The TP53 gene is a key tumor suppressor. Although the tumor suppressor p53 was one of the first to be characterized as a transcription factor, with its main function potentiated by its interaction with DNA, there are still many unresolved questions about its mechanism of action. Here, we demonstrate a novel role for p53 in the maintenance of nuclear architecture of cells. Using three-dimensional (3D) imaging and spectral karyotyping, as well as super resolution microscopy of DNA structure, we observe significant differences in 3D telomere signatures, DNA structure and DNA-poor spaces as well gains or losses of chromosomes, between normal and tumor cells with CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-deleted or wild-type TP53. Additionally, treatment with Nutlin-3 results in differences in nuclear architecture of telomeres in wild-type but not in p53 knockout MCF-7 (Michigan Cancer Foundation-7) cells. Nutlin-3 binds to the p53-binding pocket of mouse double minute 2 (MDM2) and blocks the p53-MDM2 interaction. Moreover, we demonstrate that another p53 stabilizing small molecule, RITA (reactivation of p53 and induction of tumor cell apoptosis), also induces changes in 3D DNA structure, apparently in a p53 independent manner. These results implicate p53 activity in regulating nuclear organization and, additionally, highlight the divergent effects of the p53 targeting compounds Nutlin-3 and RITA. |
format | Online Article Text |
id | pubmed-7073688 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-70736882020-03-19 p53 CRISPR Deletion Affects DNA Structure and Nuclear Architecture Rangel-Pozzo, Aline Booth, Samuel Yu, Pak Lok Ivan Singh, Madhurendra Selivanova, Galina Mai, Sabine J Clin Med Article The TP53 gene is a key tumor suppressor. Although the tumor suppressor p53 was one of the first to be characterized as a transcription factor, with its main function potentiated by its interaction with DNA, there are still many unresolved questions about its mechanism of action. Here, we demonstrate a novel role for p53 in the maintenance of nuclear architecture of cells. Using three-dimensional (3D) imaging and spectral karyotyping, as well as super resolution microscopy of DNA structure, we observe significant differences in 3D telomere signatures, DNA structure and DNA-poor spaces as well gains or losses of chromosomes, between normal and tumor cells with CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-deleted or wild-type TP53. Additionally, treatment with Nutlin-3 results in differences in nuclear architecture of telomeres in wild-type but not in p53 knockout MCF-7 (Michigan Cancer Foundation-7) cells. Nutlin-3 binds to the p53-binding pocket of mouse double minute 2 (MDM2) and blocks the p53-MDM2 interaction. Moreover, we demonstrate that another p53 stabilizing small molecule, RITA (reactivation of p53 and induction of tumor cell apoptosis), also induces changes in 3D DNA structure, apparently in a p53 independent manner. These results implicate p53 activity in regulating nuclear organization and, additionally, highlight the divergent effects of the p53 targeting compounds Nutlin-3 and RITA. MDPI 2020-02-22 /pmc/articles/PMC7073688/ /pubmed/32098416 http://dx.doi.org/10.3390/jcm9020598 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 Rangel-Pozzo, Aline Booth, Samuel Yu, Pak Lok Ivan Singh, Madhurendra Selivanova, Galina Mai, Sabine p53 CRISPR Deletion Affects DNA Structure and Nuclear Architecture |
title | p53 CRISPR Deletion Affects DNA Structure and Nuclear Architecture |
title_full | p53 CRISPR Deletion Affects DNA Structure and Nuclear Architecture |
title_fullStr | p53 CRISPR Deletion Affects DNA Structure and Nuclear Architecture |
title_full_unstemmed | p53 CRISPR Deletion Affects DNA Structure and Nuclear Architecture |
title_short | p53 CRISPR Deletion Affects DNA Structure and Nuclear Architecture |
title_sort | p53 crispr deletion affects dna structure and nuclear architecture |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7073688/ https://www.ncbi.nlm.nih.gov/pubmed/32098416 http://dx.doi.org/10.3390/jcm9020598 |
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