The Mechanism Selecting the Guide Strand from Small RNA Duplexes is Different Among Argonaute Proteins
Abstract
Double-stranded RNA induces RNA silencing and is cleaved into 21-24 nt small RNA duplexes by Dicer enzyme. A strand of Dicer-generated small RNA duplex (called the guide strand) is then selected by a thermodynamic mechanism to associate with Argonaute (AGO) protein. This AGO-small RNA complex functions to cleave mRNA, repress translation or modify chromatin structure in a sequence-specific manner. Although a model plant, Arabidopsis thaliana, contains 10 AGO genes, their roles and molecular mechanisms remain obscure. In this study, we analyzed the roles of Arabidopsis AGO2 and AGO5. Interestingly, the 5' nucleotide of small RNAs that associated with AGO2 was mainly adenine (85.7%) and that with AGO5 was mainly cytosine (83.5%). Small RNAs that were abundantly cloned from the AGO2 immunoprecipitation fraction (miR163-LL, which is derived from the Lower Left of mature miR163 in pre-miR163, and miR390) and from the AGO5 immunoprecipitation fraction (miR163-UL, which is derived from the Upper Left of mature miR163 in pre-miR163, and miR390(*)) are derived from the single small RNA duplexes, miR163-LL/miR163-UL and miR390/miR390(*). Each strand of the miR163-LL/miR163-UL duplex is selectively sorted to associate with AGO2 or AGO5 in a 5' nucleotide-dependent manner rather than in a thermodynamic stability-dependent manner. Furthermore, we showed that both AGO2 and AGO5 have the ability to bind cucumber mosaic virus-derived small RNAs. These results clearly indicate that the mechan…
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The most commonly observed 5' nucleotide of miRNA guide strands is Uracil, whereas the passenger strand often begins with Cytosine at the 5' end.
“Consistently, the most commonly observed 5′ nucleotide of miRNA guide strands is a Uracil, whereas the passenger strand often begins with a Cytosine at the 5′ end (Ghildiyal et al., 2009; Mi et al., 2008; Takeda, Iwasaki, Watanabe, Utsumi, & Watanabe, 2008; Warf, Johnson, & Bass, 2011; Figure 2c).”
From microRNA strand selection: Unwinding the rules (2021) · cites this paper as Takeda et al., 2008
While often supported by in vivo observations, how well the rules of miRNA strand selection hold up in a living organism throughout its lifespan is yet to be established.
“While often supported by in vivo observations, such as the observation that most miRNAs begin with a 5′ Uracil (Ghildiyal et al., 2009; Mi et al., 2008; Takeda et al., 2008; Warf et al., 2011), how well these rules hold up in a living organism throughout its lifespan is yet to be established.”
From microRNA strand selection: Unwinding the rules (2021) · cites this paper as Takeda et al., 2008
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- Authors
- Takeda, A., Iwasaki, S., Watanabe, T., Utsumi, M., & Watanabe, Y.
- Venue
- Plant and Cell Physiology
- Publisher
- Oxford University Press (OUP)
- Published
- 2008-04
- Volume
- 49
- Issue
- 4
- Pages
- 493-500
- ISSN
- 1471-9053
- 0032-0781
- Cited by
- 444
- References
- 0
- Language
- en
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