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 journalArticle

248 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)997-1003
Number of pages7
JournalNature Structural and Molecular Biology
Volume17
Issue number8
DOIs
StatePublished - Aug 1 2010

Fingerprint

MicroRNAs
Arabidopsis
Small Interfering RNA
Nucleotides
RNA Replicase
RNA Interference
RNA
Messenger RNA

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

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, 01.08.2010, p. 997-1003.

Research output: Contribution to journalArticle

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.
@article{ce24960faaa74fc58149c1e12cbc856f,
title = "Unique functionality of 22-nt miRNAs in triggering RDR6-dependent siRNA biogenesis from target transcripts in Arabidopsis",
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.",
author = "Cuperus, {Josh T.} and Alberto Carbonell and Noah Fahlgren and Hernan Garcia-Ruiz and Burke, {Russell T.} and Atsushi Takeda and Sullivan, {Christopher M.} and Gilbert, {Sunny D.} and Montgomery, {Taiowa A.} and Carrington, {James C.}",
year = "2010",
month = "8",
day = "1",
doi = "10.1038/nsmb.1866",
language = "English (US)",
volume = "17",
pages = "997--1003",
journal = "Nature Structural and Molecular Biology",
issn = "1545-9993",
publisher = "Nature Publishing Group",
number = "8",

}

TY - JOUR

T1 - Unique functionality of 22-nt miRNAs in triggering RDR6-dependent siRNA biogenesis from target transcripts in Arabidopsis

AU - Cuperus, Josh T.

AU - Carbonell, Alberto

AU - Fahlgren, Noah

AU - Garcia-Ruiz, Hernan

AU - Burke, Russell T.

AU - Takeda, Atsushi

AU - Sullivan, Christopher M.

AU - Gilbert, Sunny D.

AU - Montgomery, Taiowa A.

AU - Carrington, James C.

PY - 2010/8/1

Y1 - 2010/8/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=77955418697&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77955418697&partnerID=8YFLogxK

U2 - 10.1038/nsmb.1866

DO - 10.1038/nsmb.1866

M3 - Article

C2 - 20562854

AN - SCOPUS:77955418697

VL - 17

SP - 997

EP - 1003

JO - Nature Structural and Molecular Biology

JF - Nature Structural and Molecular Biology

SN - 1545-9993

IS - 8

ER -