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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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Detalles Bibliográficos
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
Descripción
Sumario: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.