In vivo imaging of tagged mRNA in plant tissues using the bacterial transcriptional antiterminator BglG.
Fiche publication
Date publication
octobre 2020
Journal
The Plant journal : for cell and molecular biology
Auteurs
Membres identifiés du Cancéropôle Est :
Dr HEINLEIN Manfred
Tous les auteurs :
Peña EJ, Robles Luna G, Heinlein M
Lien Pubmed
Résumé
RNA transport and localization represent important post-transcriptional mechanisms to determine the subcellular localization of protein synthesis. Plants have the capacity to transport mRNA molecules beyond the cell boundaries through plasmodesmata (PD) and over long distances in the phloem. RNA viruses exploit these transport pathways to disseminate their infections and represent important model systems to investigate RNA transport in plants. Here, we present an in vivo plant RNA-labeling system based on the E.coli RNA-binding protein BglG. Using the detection of RNA in mobile RNA particles formed by viral movement protein (MP) as a model, we demonstrate the efficiency and specificity of mRNA detection by the BglG system as compared to MS2 and λN systems. Our observations show that MP mRNA is specifically associated with MP in mobile MP particles but hardly with MP localized at PD. MP mRNA is clearly absent from MP accumulating along microtubules. We show that the in vivo BglG labeling of the MP particles depends on the presence of the BglG-binding stem-loop aptamers within the MP mRNA and that the aptamers also enhance the coprecipitation of BglG by MP, thus demonstrating the presence of a MP:MP mRNA complex. The BglG system also allowed us to monitor the cell-to-cell transport of the MP mRNA, thus linking the observation of mobile MP mRNA granules with intercellular MP mRNA transport. Given its specificity demonstrated here, the BglG system may be widely applicable for studying mRNA transport and localization in plants.
Mots clés
BglG, MS2, RNA imaging, RNA labeling, RNA localization, RNA transport, TMV, cell-to-cell movement, movement protein, λN
Référence
Plant J. 2020 Oct 24;: