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Promoter polymorphisms in genes involved in porcine myogenesis influence their transcriptional activity

BACKGROUND: Success of meat production and selection for improvement of meat quality is among the primary aims in animal production. Meat quality traits are economically important in swine; however, the underlying genetic nature is very complex. Therefore, an improved pork production strongly depend...

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Autores principales: Bongiorni, Silvia, Tilesi, Francesca, Bicorgna, Silvia, Iacoponi, Francesca, Willems, Daniela, Gargani, Maria, D’Andrea, MariaSilvia, Pilla, Fabio, Valentini, Alessio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4226869/
https://www.ncbi.nlm.nih.gov/pubmed/25377122
http://dx.doi.org/10.1186/s12863-014-0119-2
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author Bongiorni, Silvia
Tilesi, Francesca
Bicorgna, Silvia
Iacoponi, Francesca
Willems, Daniela
Gargani, Maria
D’Andrea, MariaSilvia
Pilla, Fabio
Valentini, Alessio
author_facet Bongiorni, Silvia
Tilesi, Francesca
Bicorgna, Silvia
Iacoponi, Francesca
Willems, Daniela
Gargani, Maria
D’Andrea, MariaSilvia
Pilla, Fabio
Valentini, Alessio
author_sort Bongiorni, Silvia
collection PubMed
description BACKGROUND: Success of meat production and selection for improvement of meat quality is among the primary aims in animal production. Meat quality traits are economically important in swine; however, the underlying genetic nature is very complex. Therefore, an improved pork production strongly depends on identifying and studying how genetic variations contribute to modulate gene expression. Promoters are key regions in gene modulation as they harbour several binding motifs to transcription regulatory factors. Therefore, polymorphisms in these regions are likely to deeply affect RNA levels and consequently protein synthesis. In this study, we report the identification of single nucleotide polymorphisms (SNPs) in promoter regions of candidate genes involved in development, cellular differentiation and muscle growth in Sus scrofa. We identified SNPs in the promoter regions of genes belonging to the Myogenic Regulatory Factors (MRF) gene family (the Myogenic Differentiation gene, MYOD1) and to Growth and Differentiation Factors (GDF) gene family (Myostatin gene, MSTN, GDF8), in Casertana and Large White breeds. The purpose of this study was to investigate if polymorphisms in the promoters could affect the transcriptional activity of these genes. With this aim, we evaluated in vitro the functional activity of the luciferase reporter gene luc2 activity, driven by two constructs carrying different promoter haplotypes. RESULTS: We tested the effects of the G302A (U12574) transition on the promoter efficiency in MYOD1 gene. We ascertained a difference in transcription efficiency for the two variants. A stronger activity of the A-carrying construct is more evident in C2C12. The luciferase expression driven by the MYOD1-A allelic variant displayed a 3.8-fold increased transcriptional activity. We investigated the activity of two haplotype variants (AY527152) in the promoter of GDF8 gene. The haploptype-1 (A435-A447-A879) up-regulated the expression of the reporter gene by a two-fold increase, and hence presumably of the GDF8 gene, in both CHO and C2C12 cultured cells. CONCLUSIONS: In vitro the MYOD1-A allelic variant could up-regulate the expression of MYOD1 gene. Additionally, we could assess a different response of in vitro gene expression according to cell type used to transfect constructs, suggesting that MyoD activation is regulated by mechanisms that are specific of myoblasts. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12863-014-0119-2) contains supplementary material, which is available to authorized users.
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spelling pubmed-42268692014-11-12 Promoter polymorphisms in genes involved in porcine myogenesis influence their transcriptional activity Bongiorni, Silvia Tilesi, Francesca Bicorgna, Silvia Iacoponi, Francesca Willems, Daniela Gargani, Maria D’Andrea, MariaSilvia Pilla, Fabio Valentini, Alessio BMC Genet Research Article BACKGROUND: Success of meat production and selection for improvement of meat quality is among the primary aims in animal production. Meat quality traits are economically important in swine; however, the underlying genetic nature is very complex. Therefore, an improved pork production strongly depends on identifying and studying how genetic variations contribute to modulate gene expression. Promoters are key regions in gene modulation as they harbour several binding motifs to transcription regulatory factors. Therefore, polymorphisms in these regions are likely to deeply affect RNA levels and consequently protein synthesis. In this study, we report the identification of single nucleotide polymorphisms (SNPs) in promoter regions of candidate genes involved in development, cellular differentiation and muscle growth in Sus scrofa. We identified SNPs in the promoter regions of genes belonging to the Myogenic Regulatory Factors (MRF) gene family (the Myogenic Differentiation gene, MYOD1) and to Growth and Differentiation Factors (GDF) gene family (Myostatin gene, MSTN, GDF8), in Casertana and Large White breeds. The purpose of this study was to investigate if polymorphisms in the promoters could affect the transcriptional activity of these genes. With this aim, we evaluated in vitro the functional activity of the luciferase reporter gene luc2 activity, driven by two constructs carrying different promoter haplotypes. RESULTS: We tested the effects of the G302A (U12574) transition on the promoter efficiency in MYOD1 gene. We ascertained a difference in transcription efficiency for the two variants. A stronger activity of the A-carrying construct is more evident in C2C12. The luciferase expression driven by the MYOD1-A allelic variant displayed a 3.8-fold increased transcriptional activity. We investigated the activity of two haplotype variants (AY527152) in the promoter of GDF8 gene. The haploptype-1 (A435-A447-A879) up-regulated the expression of the reporter gene by a two-fold increase, and hence presumably of the GDF8 gene, in both CHO and C2C12 cultured cells. CONCLUSIONS: In vitro the MYOD1-A allelic variant could up-regulate the expression of MYOD1 gene. Additionally, we could assess a different response of in vitro gene expression according to cell type used to transfect constructs, suggesting that MyoD activation is regulated by mechanisms that are specific of myoblasts. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12863-014-0119-2) contains supplementary material, which is available to authorized users. BioMed Central 2014-11-07 /pmc/articles/PMC4226869/ /pubmed/25377122 http://dx.doi.org/10.1186/s12863-014-0119-2 Text en © Bongiorni et al.; licensee BioMed Central Ltd. 2014 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 work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Bongiorni, Silvia
Tilesi, Francesca
Bicorgna, Silvia
Iacoponi, Francesca
Willems, Daniela
Gargani, Maria
D’Andrea, MariaSilvia
Pilla, Fabio
Valentini, Alessio
Promoter polymorphisms in genes involved in porcine myogenesis influence their transcriptional activity
title Promoter polymorphisms in genes involved in porcine myogenesis influence their transcriptional activity
title_full Promoter polymorphisms in genes involved in porcine myogenesis influence their transcriptional activity
title_fullStr Promoter polymorphisms in genes involved in porcine myogenesis influence their transcriptional activity
title_full_unstemmed Promoter polymorphisms in genes involved in porcine myogenesis influence their transcriptional activity
title_short Promoter polymorphisms in genes involved in porcine myogenesis influence their transcriptional activity
title_sort promoter polymorphisms in genes involved in porcine myogenesis influence their transcriptional activity
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4226869/
https://www.ncbi.nlm.nih.gov/pubmed/25377122
http://dx.doi.org/10.1186/s12863-014-0119-2
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