Argonaute (AGO) family proteins are effectors of RNAi in eukaryotes. AGOs bind small RNAs and use them as guides to silence target genes or transposable elements at the transcriptional or posttranscriptional level. Eukaryotic AGO proteins share common structural and biochemical properties and function through conserved core mechanisms in RNAi pathways, yet plant AGOs have evolved specialized and diversified functions. This Review covers the general features of AGO proteins and highlights recent progress toward our understanding of the mechanisms and functions of plant AGOs.

As key components in the eukaryotic gene regulatory network, microRNAs (miRNAs) themselves are regulated at the level of both metabolism and activity. To identify factors that modulate miRNA activity, we used an Arabidopsis thaliana transgenic line expressing an artificial miRNA that causes trichome clustering and performed a screen for mutants with compromised miRNA activity (cma mutants) or enhanced miRNA activity (ema mutants). From this screen, we identified two novel mutant alleles of SERRATE, which is known to be required for miRNA biogenesis and dozens of other cma and ema mutants. In this study, we analyzed ema1. SAD2/EMA1 encodes an Importin β protein. The ema1 mutation had no effects on the accumulation of miRNAs and ARGONAUTE1 (AGO1) or on their cytoplasmic and nuclear distributions. Intriguingly, we found that the miRNA effector complexes purified from ema1 contained a larger amount of miRNAs and displayed elevated mRNA cleavage activities, indicating that EMA1 modulates miRNA activity by influencing the loading of miRNAs into AGO1 complexes. These results implicate EMA1 as a negative regulator of the miRNA pathway and reveal a novel layer of miRNA activity modulation.

The timing of flowering is vital for plant reproductive success and is therefore tightly regulated by endogenous and exogenous cues. In summer annual Arabidopsis (Arabidopsis thaliana) accessions, like Columbia-0, rapid flowering is promoted by repression of the floral repressor FLOWERING LOCUS C (FLC). This is through the activity of the autonomous pathway, a group of proteins with diverse functions including RNA 3′-end processing factors, spliceosome components, a transcription elongation factor, and chromatin modifiers. These factors function at the FLC locus linking alternative processing of an antisense long noncoding RNA, called COOLAIR, with delivery of a repressive chromatin environment that affects the transcriptional output. The transcriptional output feeds back to influence the chromatin environment, reinforcing and stabilizing that state. This review summarizes our current knowledge of the autonomous pathway and compares it with similar cotranscriptional mechanisms in other organisms.