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The ATRX cDNA is prone to bacterial IS10 element insertions that alter its structure
The SWI/SNF-like chromatin-remodeling protein ATRX has emerged as a key factor in the regulation of α-globin gene expression, incorporation of histone variants into the chromatin template and, more recently, as a frequently mutated gene across a wide spectrum of cancers. Therefore, the availability...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer International Publishing
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4021028/ https://www.ncbi.nlm.nih.gov/pubmed/24834375 http://dx.doi.org/10.1186/2193-1801-3-222 |
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author | Valle-García, David Griffiths, Lyra M Dyer, Michael A Bernstein, Emily Recillas-Targa, Félix |
author_facet | Valle-García, David Griffiths, Lyra M Dyer, Michael A Bernstein, Emily Recillas-Targa, Félix |
author_sort | Valle-García, David |
collection | PubMed |
description | The SWI/SNF-like chromatin-remodeling protein ATRX has emerged as a key factor in the regulation of α-globin gene expression, incorporation of histone variants into the chromatin template and, more recently, as a frequently mutated gene across a wide spectrum of cancers. Therefore, the availability of a functional ATRX cDNA for expression studies is a valuable tool for the scientific community. We have identified two independent transposon insertions of a bacterial IS10 element into exon 8 of ATRX isoform 2 coding sequence in two different plasmids derived from a single source. We demonstrate that these insertion events are common and there is an insertion hotspot within the ATRX cDNA. Such IS10 insertions produce a truncated form of ATRX, which significantly compromises its nuclear localization. In turn, we describe ways to prevent IS10 insertion during propagation and cloning of ATRX-containing vectors, including optimal growth conditions, bacterial strains, and suggested sequencing strategies. Finally, we have generated an insertion-free plasmid that is available to the community for expression studies of ATRX. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/2193-1801-3-222) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4021028 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Springer International Publishing |
record_format | MEDLINE/PubMed |
spelling | pubmed-40210282014-05-15 The ATRX cDNA is prone to bacterial IS10 element insertions that alter its structure Valle-García, David Griffiths, Lyra M Dyer, Michael A Bernstein, Emily Recillas-Targa, Félix Springerplus Research The SWI/SNF-like chromatin-remodeling protein ATRX has emerged as a key factor in the regulation of α-globin gene expression, incorporation of histone variants into the chromatin template and, more recently, as a frequently mutated gene across a wide spectrum of cancers. Therefore, the availability of a functional ATRX cDNA for expression studies is a valuable tool for the scientific community. We have identified two independent transposon insertions of a bacterial IS10 element into exon 8 of ATRX isoform 2 coding sequence in two different plasmids derived from a single source. We demonstrate that these insertion events are common and there is an insertion hotspot within the ATRX cDNA. Such IS10 insertions produce a truncated form of ATRX, which significantly compromises its nuclear localization. In turn, we describe ways to prevent IS10 insertion during propagation and cloning of ATRX-containing vectors, including optimal growth conditions, bacterial strains, and suggested sequencing strategies. Finally, we have generated an insertion-free plasmid that is available to the community for expression studies of ATRX. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/2193-1801-3-222) contains supplementary material, which is available to authorized users. Springer International Publishing 2014-05-02 /pmc/articles/PMC4021028/ /pubmed/24834375 http://dx.doi.org/10.1186/2193-1801-3-222 Text en © Valle-García et al.; licensee Springer. 2014 This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. |
spellingShingle | Research Valle-García, David Griffiths, Lyra M Dyer, Michael A Bernstein, Emily Recillas-Targa, Félix The ATRX cDNA is prone to bacterial IS10 element insertions that alter its structure |
title | The ATRX cDNA is prone to bacterial IS10 element insertions that alter its structure |
title_full | The ATRX cDNA is prone to bacterial IS10 element insertions that alter its structure |
title_fullStr | The ATRX cDNA is prone to bacterial IS10 element insertions that alter its structure |
title_full_unstemmed | The ATRX cDNA is prone to bacterial IS10 element insertions that alter its structure |
title_short | The ATRX cDNA is prone to bacterial IS10 element insertions that alter its structure |
title_sort | atrx cdna is prone to bacterial is10 element insertions that alter its structure |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4021028/ https://www.ncbi.nlm.nih.gov/pubmed/24834375 http://dx.doi.org/10.1186/2193-1801-3-222 |
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