Article abstract
Nature Structural & Molecular Biology 14, 927 - 933 (2007)
Published online: 23 September 2007 | doi:10.1038/nsmb1308
Structural features of small RNA precursors determine Argonaute loading in Caenorhabditis elegans
Florian A Steiner1, Suzanne W Hoogstrate1, Kristy L Okihara1, Karen L Thijssen1,2, Rene F Ketting1, Ronald H A Plasterk1 & Titia Sijen1,2
Abstract
In C. elegans, DCR-1 is required for the maturation of both short interfering RNAs (siRNAs) and microRNAs (miRNAs), which are subsequently loaded into different Argonaute proteins to mediate silencing via distinct mechanisms. We used in vivo analyses to show that precursors of small RNAs contain structural features that direct the small RNAs into the RNA interference (RNAi) pathway or the miRNA-processing pathway. Nucleotide changes in the pre-let-7 miRNA precursor that make its stem fully complementary cause the resulting small RNA to be recognized as siRNA and induce binding to RDE-1, which leads to RNAi. Mismatches of 1 to 3 nucleotides at various positions in the stem of the precursor restore direction into the miRNA pathway, as the largest portion of such small RNA variants is associated with ALG-1. The Argonaute proteins to which the small RNAs are bound determine the silencing mode, and no functional overlap between RDE-1 and ALG-1 was detected.
- Hubrecht Institute, Koninklijke Nederlandse Akademie van Wetenschappen, Utrecht, 3584 CT, The Netherlands.
- Present addresses: University Medical Center Groningen, Groningen, 9700 RB, The Netherlands (K.L.T.) and Netherlands Forensic Institute, The Hague, 2497 GB, The Netherlands (T.S.).
Correspondence to: Titia Sijen1,2 e-mail: t.sijen@nfi.minjus.nl
Correspondence to: Florian A Steiner1 e-mail: f.steiner@niob.knaw.nl
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