Unique functionality of 22-nt miRNAs in triggering RDR6-dependent siRNA biogenesis from target transcripts in Arabidopsis

Josh T. Cuperus; Alberto Carbonell; Noah Fahlgren; Hernan Garcia-Ruiz; Russell T. Burke; Atsushi Takeda; Christopher M. Sullivan; Sunny D. Gilbert; Taiowa A. Montgomery; James C. Carrington

doi:10.1038/nsmb.1866

Unique functionality of 22-nt miRNAs in triggering RDR6-dependent siRNA biogenesis from target transcripts in Arabidopsis

Josh T. Cuperus, Alberto Carbonell, Noah Fahlgren, Hernan Garcia-Ruiz, Russell T. Burke, Atsushi Takeda, Christopher M. Sullivan, Sunny D. Gilbert, Taiowa A. Montgomery, James C. Carrington

Research output: Contribution to journal › Article › peer-review

267 Scopus citations

Abstract

RNA interference pathways can involve amplification of secondary siRNAs by RNA-dependent RNA polymerases. In plants, RDR6-dependent secondary siRNAs arise from transcripts targeted by some microRNAs (miRNAs). Here, Arabidopsis thaliana secondary siRNAs from mRNA as well as trans-acting siRNAs are shown to be triggered through initial targeting by a 22-nucleotide (nt) miRNA that associates with AGO1. In contrast to canonical 21-nt miRNAs, 22-nt miRNAs primarily arise from foldback precursors containing asymmetric bulges. Using artificial miRNA constructs, conversion of asymmetric foldbacks to symmetric foldbacks resulted in the production of 21-nt forms of miR173, miR472 and miR828. Both 21-and 22-nt forms associated with AGO1 and guided accurate slicer activity, but only 22-nt forms were competent to trigger RDR6-dependent siRNA production from target RNA. These data suggest that AGO1 functions differentially with 21-and 22-nt miRNAs to engage the RDR6-associated amplification apparatus.

Original language	English (US)
Pages (from-to)	997-1003
Number of pages	7
Journal	Nature Structural and Molecular Biology
Volume	17
Issue number	8
DOIs	https://doi.org/10.1038/nsmb.1866
State	Published - Aug 2010
Externally published	Yes

ASJC Scopus subject areas

Structural Biology
Molecular Biology

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Cuperus, J. T., Carbonell, A., Fahlgren, N., Garcia-Ruiz, H., Burke, R. T., Takeda, A., Sullivan, C. M., Gilbert, S. D., Montgomery, T. A., & Carrington, J. C. (2010). Unique functionality of 22-nt miRNAs in triggering RDR6-dependent siRNA biogenesis from target transcripts in Arabidopsis. Nature Structural and Molecular Biology, 17(8), 997-1003. https://doi.org/10.1038/nsmb.1866

Unique functionality of 22-nt miRNAs in triggering RDR6-dependent siRNA biogenesis from target transcripts in Arabidopsis. / Cuperus, Josh T.; Carbonell, Alberto; Fahlgren, Noah; Garcia-Ruiz, Hernan; Burke, Russell T.; Takeda, Atsushi; Sullivan, Christopher M.; Gilbert, Sunny D.; Montgomery, Taiowa A.; Carrington, James C.

In: Nature Structural and Molecular Biology, Vol. 17, No. 8, 08.2010, p. 997-1003.

Research output: Contribution to journal › Article › peer-review

Cuperus, JT, Carbonell, A, Fahlgren, N, Garcia-Ruiz, H, Burke, RT, Takeda, A, Sullivan, CM, Gilbert, SD, Montgomery, TA & Carrington, JC 2010, 'Unique functionality of 22-nt miRNAs in triggering RDR6-dependent siRNA biogenesis from target transcripts in Arabidopsis', Nature Structural and Molecular Biology, vol. 17, no. 8, pp. 997-1003. https://doi.org/10.1038/nsmb.1866

Cuperus, Josh T. ; Carbonell, Alberto ; Fahlgren, Noah ; Garcia-Ruiz, Hernan ; Burke, Russell T. ; Takeda, Atsushi ; Sullivan, Christopher M. ; Gilbert, Sunny D. ; Montgomery, Taiowa A. ; Carrington, James C. / Unique functionality of 22-nt miRNAs in triggering RDR6-dependent siRNA biogenesis from target transcripts in Arabidopsis. In: Nature Structural and Molecular Biology. 2010 ; Vol. 17, No. 8. pp. 997-1003.

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abstract = "RNA interference pathways can involve amplification of secondary siRNAs by RNA-dependent RNA polymerases. In plants, RDR6-dependent secondary siRNAs arise from transcripts targeted by some microRNAs (miRNAs). Here, Arabidopsis thaliana secondary siRNAs from mRNA as well as trans-acting siRNAs are shown to be triggered through initial targeting by a 22-nucleotide (nt) miRNA that associates with AGO1. In contrast to canonical 21-nt miRNAs, 22-nt miRNAs primarily arise from foldback precursors containing asymmetric bulges. Using artificial miRNA constructs, conversion of asymmetric foldbacks to symmetric foldbacks resulted in the production of 21-nt forms of miR173, miR472 and miR828. Both 21-and 22-nt forms associated with AGO1 and guided accurate slicer activity, but only 22-nt forms were competent to trigger RDR6-dependent siRNA production from target RNA. These data suggest that AGO1 functions differentially with 21-and 22-nt miRNAs to engage the RDR6-associated amplification apparatus.",

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