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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...

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Autores principales: Valle-García, David, Griffiths, Lyra M, Dyer, Michael A, Bernstein, Emily, Recillas-Targa, Félix
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer International Publishing 2014
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.
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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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