Transcriptomics,Genomics

Dataset Information

99

Arabidopsis thaliana AGO1 immunoprecipitated small RNA from Col0 plants transformed with a transgene expressing wild-type or a nuclear localized form of AGO1 fused to GFP.


ABSTRACT: Plant microRNAs undergo stepwise-nuclear maturation before engaging cytosolic sequence-complementary transcripts in association with the silencing-effector protein ARGONAUTE1(AGO1). How plant miRNAs translocate to the cytosol remains mysterious as does their cellular loading site(s) into AGO1. Here, we show that the N-termini of plant AGO1s contain a nuclear-localization (NLS) and nuclear-export signal (NES), which, in Arabidopsis thaliana (At) enables AtAGO1 nucleo-cytosolic shuttling in a Leptomycin-B-inhibited manner, diagnostic of CRM1(Expo1)/NES-dependent nuclear export. Nuclear-only AtAGO1 contains the same 2’O-methylated miRNA cohort as its nucleo-cytosolic counterpart, but specifically interacts with the miRNA loading-chaperone HSP90. AtAGO1 nucleo-cytosolic shuttling is required for mature miRNA translocation and for miRNA-mediated silencing. We propose that plant miRNAs are matured, methylated, loaded into AGO1 in the nucleus, and exported cytoplasmically as AGO1:miRNA complexes in a CRM1(Expo1)/NES-dependent manner. Overall design: 2 samples examined: Arabidopsis thaliana AGO1 immunoprecipitated small RNA Col0 from plants expressing pAGO1:GFP-AGO1 or pAGO1:GFP-AGO1ΔNES (2 replicates for each)

INSTRUMENT(S): Illumina HiSeq 2500 (Arabidopsis thaliana)

SUBMITTER: Alexis Sarazin  

PROVIDER: GSE100395 | GEO | 2018-01-04

SECONDARY ACCESSION(S): PRJNA391518

REPOSITORIES: GEO

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Publications

Nucleo-cytosolic Shuttling of ARGONAUTE1 Prompts a Revised Model of the Plant MicroRNA Pathway.

Bologna Nicolas G NG   Iselin Raphael R   Abriata Luciano A LA   Sarazin Alexis A   Pumplin Nathan N   Jay Florence F   Grentzinger Thomas T   Dal Peraro Matteo M   Voinnet Olivier O  

Molecular cell 20180201 4


Unlike in metazoans, plant microRNAs (miRNAs) undergo stepwise nuclear maturation before engaging cytosolic, sequence-complementary transcripts in association with the silencing effector protein ARGONAUTE1 (AGO1). Since their discovery, how and under which form plant miRNAs translocate to the cytosol has remained unclear, as has their sub-cellular AGO1 loading site(s). Here, we show that the N termini of all plant AGO1s contain a nuclear-localization (NLS) and nuclear-export signal (NES) that, i  ...[more]

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