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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 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.

SUBMITTER: Hameed MW 

PROVIDER: S-EPMC5729415 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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

Hameed Muhammad Waqar MW   Juszczak Ilona I   Bock Ralph R   van Dongen Joost Thomas JT  

Plant methods 20171213


<h4>Background</h4>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 <i>Nicotiana tabacum</i> and <i>Arabidopsis thaliana</i> mitochondrial mRNA.<h4>Results</h4>Microarray analysis of free and polysomal mRNAs was used to c  ...[more]

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