R2D2 organizes small regulatory RNA pathways in Drosophila Journal Article
| Authors: | Okamura, K.; Robine, N.; Liu, Y.; Liu, Q.; Lai, E. C. |
| Article Title: | R2D2 organizes small regulatory RNA pathways in Drosophila |
| Abstract: | Drosophila microRNAs (miRNAs) and small interfering RNAs (siRNAs) are generally produced by different Dicer enzymes (Dcr-1 and Dcr-2) and sorted to functionally distinct Argonaute effectors (AGO1 and AGO2). However, there is cross talk between these pathways, as highlighted by the recognition that Drosophila miRNA* strands (the partner strands of mature miRNAs) are generated by Dcr-1 but are preferentially sorted to AGO2. Here, we show that a component of the siRNA loading complex, R2D2, is essential both to load endogenously encoded siRNAs (endo-siRNAs) into AGO2 and to prevent endosiRNAs from binding to AGO1. Northern blot analysis and deep sequencing showed that in the r2d2 mutant, all classes of endo-siRNAs were unable to load AGO2 and instead accumulated in the AGO1 complex. Such redirection was specific to endo-siRNAs and was not observed with miRNA* strands. We observed functional consequences of altered sorting in RNA interference (RNAi) mutants, since endosiRNAs generated from cis-natural antisense transcripts (cis-NAT-siRNA) exhibited evidence for biased maturation as single strands in AGO1 according to thermodynamic asymmetry and a hairpin-derived endo-siRNA formed cleavage-competent complexes with AGO1 upon mutation of r2d2. Finally, we demonstrated a direct role for the R2D2/Dcr-2 heterodimer in sensing central mismatch positions that direct miRNA* strands to AGO2. Together, these data reveal new roles of R2D2 in organizing small RNA networks in Drosophila. Copyright © 2011, American Society for Microbiology. All Rights Reserved. |
| Keywords: | unclassified drug; gene mutation; nonhuman; microrna; embryo; animal experiment; small interfering rna; drosophila; rna interference; in vitro study; rna binding protein; nucleotide sequence; gene control; thermodynamics; argonaute 1 protein; argonaute 2 protein; protein rna binding; dcr 2 enzyme; protein r2d2 |
| Journal Title: | Molecular and Cellular Biology |
| Volume: | 31 |
| Issue: | 4 |
| ISSN: | 0270-7306 |
| Publisher: | American Society for Microbiology |
| Date Published: | 2011-02-01 |
| Start Page: | 884 |
| End Page: | 896 |
| Language: | English |
| DOI: | 10.1128/mcb.01141-10 |
| PROVIDER: | scopus |
| PMCID: | PMC3028645 |
| PUBMED: | 21135122 |
| DOI/URL: | |
| Notes: | --- - "Export Date: 4 March 2011" - "CODEN: MCEBD" - "Source: Scopus" |
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