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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...

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Autores principales: Rangel-Pozzo, Aline, Booth, Samuel, Yu, Pak Lok Ivan, Singh, Madhurendra, Selivanova, Galina, Mai, Sabine
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
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.
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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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