ABSTRACT
ARGONAUTE (AGO) proteins are the final effectors of small RNA-mediated transcriptional and post-transcriptional silencing pathways. Plant AGO proteins are essential for preserving genome integrity, regulating developmental processes, and in stress responses and pathogen defense. Since the discovery of the first eukaryotic AGO in Arabidopsis, our understanding of these proteins has grown exponentially throughout all the eukaryotes. However, many aspects of AGO proteins' modes of action and how they are influenced by their subcellular localization are still to be elucidated. Here, we provide an updated and comprehensive view of the evolution, domain architecture and roles, expression pattern, subcellular localization, and biological functions of the 10 AGO proteins in Arabidopsis.
Subject(s)
Arabidopsis , MicroRNAs , Arabidopsis/genetics , Arabidopsis/metabolism , Argonaute Proteins/genetics , Argonaute Proteins/metabolism , Plant Proteins/metabolism , Plants/metabolism , MicroRNAs/metabolism , RNA InterferenceABSTRACT
The nature of plant tissues has continuously hampered understanding of the spatio-temporal and subcellular distribution of RNA-guided processes. Here, we describe a universal protocol based on Arabidopsis to investigate subcellular RNA distribution from virtually any plant species using flow cytometry sorting. This protocol includes all necessary control steps to assess the quality of the nuclear RNA purification. Moreover, it can be easily applied to different plant developmental stages, tissues, cell cycle phases, experimental growth conditions, and specific cell type(s). For complete information on the use and execution of this protocol, please refer to Bologna et al. (2018) and de Leone et al. (2020).
