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

Abstract 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 base...

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Main Authors: Muhammad Waqar Hameed, Ilona Juszczak, Ralph Bock, Joost Thomas van Dongen
Format: Article
Language:English
Published: BMC 2017-12-01
Series:Plant Methods
Subjects:
Online Access:http://link.springer.com/article/10.1186/s13007-017-0257-4
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author Muhammad Waqar Hameed
Ilona Juszczak
Ralph Bock
Joost Thomas van Dongen
author_facet Muhammad Waqar Hameed
Ilona Juszczak
Ralph Bock
Joost Thomas van Dongen
author_sort Muhammad Waqar Hameed
collection DOAJ
description Abstract 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.
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spelling doaj.art-5ea821510561406aa97b4e1368fd8e202022-12-22T00:54:18ZengBMCPlant Methods1746-48112017-12-0113111810.1186/s13007-017-0257-4Comparison of mitochondrial gene expression and polysome loading in different tobacco tissuesMuhammad Waqar Hameed0Ilona Juszczak1Ralph Bock2Joost Thomas van Dongen3Max Planck Institute of Molecular Plant PhysiologyMax Planck Institute of Molecular Plant PhysiologyMax Planck Institute of Molecular Plant PhysiologyMax Planck Institute of Molecular Plant PhysiologyAbstract 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.http://link.springer.com/article/10.1186/s13007-017-0257-4Plant mitochondriaTranscriptionTranslationMitochondrial microarrayMitochondrial ribosomes
spellingShingle Muhammad Waqar Hameed
Ilona Juszczak
Ralph Bock
Joost Thomas van Dongen
Comparison of mitochondrial gene expression and polysome loading in different tobacco tissues
Plant Methods
Plant mitochondria
Transcription
Translation
Mitochondrial microarray
Mitochondrial ribosomes
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 Plant mitochondria
Transcription
Translation
Mitochondrial microarray
Mitochondrial ribosomes
url http://link.springer.com/article/10.1186/s13007-017-0257-4
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AT ralphbock comparisonofmitochondrialgeneexpressionandpolysomeloadingindifferenttobaccotissues
AT joostthomasvandongen comparisonofmitochondrialgeneexpressionandpolysomeloadingindifferenttobaccotissues