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Nucleation, propagation and cleavage of target RNAs in Ago silencing complexes
Open accessAbstract
The slicer activity of the RNA-induced silencing complex resides within its Argonaute (Ago) component, in which the PIWI domain provides the catalytic residues governing guide-strand mediated site-specific cleavage of target RNA. Here we report on structures of ternary complexes of Thermus thermophilus Ago catalytic mutants with 5′-phosphorylated 21-nucleotide guide DNA and complementary target RNAs of 12, 15 and 19 nucleotides in length, which define the molecular basis for Mg2+-facilitated site-specific cleavage of the target. We observe pivot-like domain movements within the Ago scaffold on proceeding from nucleation to propagation steps of guide–target duplex formation, with duplex zippering beyond one turn of the helix requiring the release of the 3′-end of the guide from the PAZ pocket. Cleavage assays on targets of various lengths supported this model, and sugar-phosphate-backbone-modified target strands showed the importance of structural and catalytic divalent metal ions observed in the crystal structures.
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Metadata
- Authors
- Wang, Y., Juranek, S., Li, H., Sheng, G., Wardle, G. S., Tuschl, T., & Patel, D. J.
- Venue
- Nature
- Published
- 2009-10
- Volume
- 461
- Issue
- 7265
- Pages
- 754-761
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- OpenAlex
- W2034394760
- Cited by
- 542
- References
- 8
- FWCI
- 18.36
- Type
- journal article
- Language
- en
- Open access
- green
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- Wang, Y., Juranek, S., Li, H., Sheng, G., Wardle, G. S., Tuschl, T., & Patel, D. J. (2009). Nucleation, propagation and cleavage of target RNAs in Ago silencing complexes. Nature. https://doi.org/10.1038/nature08434
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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