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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Format: | Article |
Language: | English |
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BMC
2017-12-01
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Series: | Plant Methods |
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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. |
first_indexed | 2024-12-11T18:50:21Z |
format | Article |
id | doaj.art-5ea821510561406aa97b4e1368fd8e20 |
institution | Directory Open Access Journal |
issn | 1746-4811 |
language | English |
last_indexed | 2024-12-11T18:50:21Z |
publishDate | 2017-12-01 |
publisher | BMC |
record_format | Article |
series | Plant Methods |
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 |
work_keys_str_mv | AT muhammadwaqarhameed comparisonofmitochondrialgeneexpressionandpolysomeloadingindifferenttobaccotissues AT ilonajuszczak comparisonofmitochondrialgeneexpressionandpolysomeloadingindifferenttobaccotissues AT ralphbock comparisonofmitochondrialgeneexpressionandpolysomeloadingindifferenttobaccotissues AT joostthomasvandongen comparisonofmitochondrialgeneexpressionandpolysomeloadingindifferenttobaccotissues |