Nucleation, propagation and cleavage of target RNAs in Ago silencing complexes Journal Article
| Authors: | Wang, Y.; Juranek, S.; Li, H.; Sheng, G.; Wardle, G. S.; Tuschl, T.; Patel, D. J. |
| Article Title: | Nucleation, propagation and cleavage of target RNAs in Ago silencing complexes |
| Abstract: | 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 Mg 2+-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. © 2009 Macmillan Publishers Limited. All rights reserved. |
| Keywords: | nonhuman; chemical analysis; gene targeting; protein; structure-activity relationship; phosphorylation; rna; dna; substrate specificity; base pairing; crystal structure; models, molecular; crystallography, x-ray; gene silencing; catalysis; protein structure; biochemistry; catalytic domain; argonaute protein; rna-induced silencing complex; phosphate; rna cleavage; biocatalysis; rna sequence; magnesium ion; cation; nucleation; sugar; thermus thermophilus; cations, divalent; magnesium |
| Journal Title: | Nature |
| Volume: | 461 |
| Issue: | 7265 |
| ISSN: | 0028-0836 |
| Publisher: | Nature Publishing Group |
| Date Published: | 2009-10-08 |
| Start Page: | 754 |
| End Page: | 761 |
| Language: | English |
| DOI: | 10.1038/nature08434 |
| PUBMED: | 19812667 |
| PROVIDER: | scopus |
| PMCID: | PMC2880917 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 28" - "Export Date: 30 November 2010" - "CODEN: NATUA" - "Source: Scopus" |
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