Cargando…

A Leader Intron of a Soybean Elongation Factor 1A (eEF1A) Gene Interacts with Proximal Promoter Elements to Regulate Gene Expression in Synthetic Promoters

Introns, especially the first intron in the 5’ untranslated region (5’UTR), can significantly impact gene expression via intron-mediated enhancement (IME). In this study, we demonstrate the leader intron of a soybean elongation factor 1A (eEF1A) gene (GmScreamM8) was essential for the high activity...

Descripción completa

Detalles Bibliográficos
Autores principales: Zhang, Ning, McHale, Leah K., Finer, John J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5091777/
https://www.ncbi.nlm.nih.gov/pubmed/27806110
http://dx.doi.org/10.1371/journal.pone.0166074
_version_ 1782464633504792576
author Zhang, Ning
McHale, Leah K.
Finer, John J.
author_facet Zhang, Ning
McHale, Leah K.
Finer, John J.
author_sort Zhang, Ning
collection PubMed
description Introns, especially the first intron in the 5’ untranslated region (5’UTR), can significantly impact gene expression via intron-mediated enhancement (IME). In this study, we demonstrate the leader intron of a soybean elongation factor 1A (eEF1A) gene (GmScreamM8) was essential for the high activity of the native promoter. Furthermore, the interaction of the GmScreamM8 leader intron with regulatory element sequences from several soybean eEF1A promoters was studied using synthetic promoters, which consisted of element tetramers upstream of a core promoter used to regulate a green fluorescent protein (gfp) reporter gene. Element tetramers, placed upstream of a GmScreamM8 core promoter, showed very high activity using both transient expression in lima bean cotyledons and stable expression in soybean hairy roots, only if the native leader intron was included, suggesting an interaction between intronic sequences and promoter elements. Partial deletions of the leader intron showed that a 222 bp intronic sequence significantly contributed to very high levels of GFP expression. Generation of synthetic intron variants with a monomeric or trimeric repeat of the 222 bp intronic sequence, yielded almost two-fold higher expression compared to the original intron, while partial deletion of the 222 bp intronic repeated sequence significantly decreased gene expression, indicating that this intronic sequence was essential for the intron-element interaction enhancement.
format Online
Article
Text
id pubmed-5091777
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-50917772016-11-15 A Leader Intron of a Soybean Elongation Factor 1A (eEF1A) Gene Interacts with Proximal Promoter Elements to Regulate Gene Expression in Synthetic Promoters Zhang, Ning McHale, Leah K. Finer, John J. PLoS One Research Article Introns, especially the first intron in the 5’ untranslated region (5’UTR), can significantly impact gene expression via intron-mediated enhancement (IME). In this study, we demonstrate the leader intron of a soybean elongation factor 1A (eEF1A) gene (GmScreamM8) was essential for the high activity of the native promoter. Furthermore, the interaction of the GmScreamM8 leader intron with regulatory element sequences from several soybean eEF1A promoters was studied using synthetic promoters, which consisted of element tetramers upstream of a core promoter used to regulate a green fluorescent protein (gfp) reporter gene. Element tetramers, placed upstream of a GmScreamM8 core promoter, showed very high activity using both transient expression in lima bean cotyledons and stable expression in soybean hairy roots, only if the native leader intron was included, suggesting an interaction between intronic sequences and promoter elements. Partial deletions of the leader intron showed that a 222 bp intronic sequence significantly contributed to very high levels of GFP expression. Generation of synthetic intron variants with a monomeric or trimeric repeat of the 222 bp intronic sequence, yielded almost two-fold higher expression compared to the original intron, while partial deletion of the 222 bp intronic repeated sequence significantly decreased gene expression, indicating that this intronic sequence was essential for the intron-element interaction enhancement. Public Library of Science 2016-11-02 /pmc/articles/PMC5091777/ /pubmed/27806110 http://dx.doi.org/10.1371/journal.pone.0166074 Text en © 2016 Zhang et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Zhang, Ning
McHale, Leah K.
Finer, John J.
A Leader Intron of a Soybean Elongation Factor 1A (eEF1A) Gene Interacts with Proximal Promoter Elements to Regulate Gene Expression in Synthetic Promoters
title A Leader Intron of a Soybean Elongation Factor 1A (eEF1A) Gene Interacts with Proximal Promoter Elements to Regulate Gene Expression in Synthetic Promoters
title_full A Leader Intron of a Soybean Elongation Factor 1A (eEF1A) Gene Interacts with Proximal Promoter Elements to Regulate Gene Expression in Synthetic Promoters
title_fullStr A Leader Intron of a Soybean Elongation Factor 1A (eEF1A) Gene Interacts with Proximal Promoter Elements to Regulate Gene Expression in Synthetic Promoters
title_full_unstemmed A Leader Intron of a Soybean Elongation Factor 1A (eEF1A) Gene Interacts with Proximal Promoter Elements to Regulate Gene Expression in Synthetic Promoters
title_short A Leader Intron of a Soybean Elongation Factor 1A (eEF1A) Gene Interacts with Proximal Promoter Elements to Regulate Gene Expression in Synthetic Promoters
title_sort leader intron of a soybean elongation factor 1a (eef1a) gene interacts with proximal promoter elements to regulate gene expression in synthetic promoters
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5091777/
https://www.ncbi.nlm.nih.gov/pubmed/27806110
http://dx.doi.org/10.1371/journal.pone.0166074
work_keys_str_mv AT zhangning aleaderintronofasoybeanelongationfactor1aeef1ageneinteractswithproximalpromoterelementstoregulategeneexpressioninsyntheticpromoters
AT mchaleleahk aleaderintronofasoybeanelongationfactor1aeef1ageneinteractswithproximalpromoterelementstoregulategeneexpressioninsyntheticpromoters
AT finerjohnj aleaderintronofasoybeanelongationfactor1aeef1ageneinteractswithproximalpromoterelementstoregulategeneexpressioninsyntheticpromoters
AT zhangning leaderintronofasoybeanelongationfactor1aeef1ageneinteractswithproximalpromoterelementstoregulategeneexpressioninsyntheticpromoters
AT mchaleleahk leaderintronofasoybeanelongationfactor1aeef1ageneinteractswithproximalpromoterelementstoregulategeneexpressioninsyntheticpromoters
AT finerjohnj leaderintronofasoybeanelongationfactor1aeef1ageneinteractswithproximalpromoterelementstoregulategeneexpressioninsyntheticpromoters