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Comparison of mitochondrial gene expression and polysome loading in different tobacco tissues

BACKGROUND: To investigate translational regulation of gene expression in plant mitochondria, a mitochondrial polysome isolation protocol was established for tobacco to investigate polysomal mRNA loading as a proxy for translational activity. Furthermore, we developed an oligonucleotide based microa...

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Autores principales: Hameed, Muhammad Waqar, Juszczak, Ilona, Bock, Ralph, van Dongen, Joost Thomas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5729415/
https://www.ncbi.nlm.nih.gov/pubmed/29255478
http://dx.doi.org/10.1186/s13007-017-0257-4
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author Hameed, Muhammad Waqar
Juszczak, Ilona
Bock, Ralph
van Dongen, Joost Thomas
author_facet Hameed, Muhammad Waqar
Juszczak, Ilona
Bock, Ralph
van Dongen, Joost Thomas
author_sort Hameed, Muhammad Waqar
collection PubMed
description BACKGROUND: To investigate translational regulation of gene expression in plant mitochondria, a mitochondrial polysome isolation protocol was established for tobacco to investigate polysomal mRNA loading as a proxy for translational activity. Furthermore, we developed an oligonucleotide based microarray platform to determine the level of Nicotiana tabacum and Arabidopsis thaliana mitochondrial mRNA. RESULTS: Microarray analysis of free and polysomal mRNAs was used to characterize differences in the levels of free transcripts and ribosome-bound mRNAs in various organs of tobacco plants. We have observed higher mitochondrial transcript levels in young leaves, flowers and floral buds as compared to fully expanded leaves and roots. A similar pattern of abundance was observed for ribosome-bound mitochondrial mRNAs in these tissues. However, the accumulation of the mitochondrial protein COX2 was found to be inversely related to that of its ribosome-bound mRNA. CONCLUSIONS: Our results indicate that the association of mitochondrial mRNAs to ribosomes is largely determined by the total transcript level of a gene. However, at least for Cox2, we demonstrated that the level of ribosome-bound mRNA is not reflected by the amount of COX2 protein. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13007-017-0257-4) contains supplementary material, which is available to authorized users.
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spelling pubmed-57294152017-12-18 Comparison of mitochondrial gene expression and polysome loading in different tobacco tissues Hameed, Muhammad Waqar Juszczak, Ilona Bock, Ralph van Dongen, Joost Thomas Plant Methods Methodology BACKGROUND: To investigate translational regulation of gene expression in plant mitochondria, a mitochondrial polysome isolation protocol was established for tobacco to investigate polysomal mRNA loading as a proxy for translational activity. Furthermore, we developed an oligonucleotide based microarray platform to determine the level of Nicotiana tabacum and Arabidopsis thaliana mitochondrial mRNA. RESULTS: Microarray analysis of free and polysomal mRNAs was used to characterize differences in the levels of free transcripts and ribosome-bound mRNAs in various organs of tobacco plants. We have observed higher mitochondrial transcript levels in young leaves, flowers and floral buds as compared to fully expanded leaves and roots. A similar pattern of abundance was observed for ribosome-bound mitochondrial mRNAs in these tissues. However, the accumulation of the mitochondrial protein COX2 was found to be inversely related to that of its ribosome-bound mRNA. CONCLUSIONS: Our results indicate that the association of mitochondrial mRNAs to ribosomes is largely determined by the total transcript level of a gene. However, at least for Cox2, we demonstrated that the level of ribosome-bound mRNA is not reflected by the amount of COX2 protein. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13007-017-0257-4) contains supplementary material, which is available to authorized users. BioMed Central 2017-12-13 /pmc/articles/PMC5729415/ /pubmed/29255478 http://dx.doi.org/10.1186/s13007-017-0257-4 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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 Methodology
Hameed, Muhammad Waqar
Juszczak, Ilona
Bock, Ralph
van Dongen, Joost Thomas
Comparison of mitochondrial gene expression and polysome loading in different tobacco tissues
title Comparison of mitochondrial gene expression and polysome loading in different tobacco tissues
title_full Comparison of mitochondrial gene expression and polysome loading in different tobacco tissues
title_fullStr Comparison of mitochondrial gene expression and polysome loading in different tobacco tissues
title_full_unstemmed Comparison of mitochondrial gene expression and polysome loading in different tobacco tissues
title_short Comparison of mitochondrial gene expression and polysome loading in different tobacco tissues
title_sort comparison of mitochondrial gene expression and polysome loading in different tobacco tissues
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5729415/
https://www.ncbi.nlm.nih.gov/pubmed/29255478
http://dx.doi.org/10.1186/s13007-017-0257-4
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