Background AGO1 associates with microRNAs (miRNAs) and regulates mRNAs through cleavage and translational repression. AGO1 homeostasis entails DCL1-dependent production of miR168 from MIR168a and MIR168b transcripts, post-transcriptional stabilization of miR168 by AGO1, and AGO1-catalyzed miR168-guided cleavage of AGO1 mRNA. Principal Findings: This study reveals that MIR168a is highly expressed and predominantly produces a 21-nt miR168 species. By contrast, MIR168b is expressed at low levels and produces an equal amount of 21- and 22-nt miR168 species. Only the 21-nt miR168 is preferentially stabilized by AGO1, and consequently, the accumulation of the 22-nt but not the 21-nt miR168 is reduced when DCL1 activity is impaired. mir168a mutants with strongly reduced levels of 21-nt miR168 are viable but exhibit developmental defects, particularly during environmentally challenging conditions. Conclusions/Significance: These results suggest that 22-nt miR168 ensures basal cleavage of AGO1 mRNA whereas 21-nt miR168 permits an effective response to endogenous or environmental fluctuations owing to its preferential stabilization by AGO1.
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