Crystal structure of A. aeolicus argonaute, a site-specific DNA-guided endoribonuclease, provides insights into RISC-mediated mRNA cleavage Journal Article
| Authors: | Yuan, Y. R.; Pei, Y.; Ma, J. B. ; Kuryavyi, V.; Zhadina, M.; Meister, G.; Chen, H. Y.; Dauter, Z.; Tuschl, T.; Patel, D. J. |
| Article Title: | Crystal structure of A. aeolicus argonaute, a site-specific DNA-guided endoribonuclease, provides insights into RISC-mediated mRNA cleavage |
| Abstract: | Argonaute (Ago) proteins constitute a key component of the RNA-induced silencing complex (RISC). We report the crystal structure of Aquifex aeolicus Ago (Aa-Ago) together with binding and cleavage studies, which establish this eubacterial Ago as a bona fide guide DNA strand-mediated site-specific RNA endonuclease. We have generated a stereochemically robust model of the complex, where the guide DNA-mRNA duplex is positioned within a basic channel spanning the bilobal interface, such that the 5′ phosphate of the guide strand can be anchored in a basic pocket, and the mRNA can be positioned for site-specific cleavage by RNase H-type divalent cation-coordinated catalytic Asp residues of the PIWI domain. Domain swap experiments involving chimeras of human Ago (hAgo1) and cleavage-competent hAgo2 reinforce the role of the PIWI domain in "slicer" activity. We propose a four-step Ago-mediated catalytic cleavage cycle model, which provides distinct perspectives into the mechanism of guide strand-mediated mRNA cleavage within the RISC. Copyright ©2005 by Elsevier Inc. |
| Keywords: | controlled study; unclassified drug; human cell; nonhuman; protein conformation; protein domain; protein degradation; protein; protein binding; chimera; bacterial proteins; double stranded dna; amino acid sequence; molecular sequence data; sequence homology, amino acid; recombinant fusion proteins; messenger rna; rna, messenger; recombinant proteins; crystal structure; models, molecular; crystallography, x-ray; protein structure, tertiary; binding sites; gene silencing; dna, single-stranded; catalytic domain; argonaute protein; rna-induced silencing complex; models, chemical; rna cleavage; eukaryotic initiation factors; rna, double-stranded; oligonucleotides; enzyme active site; deoxyribonuclease; cations, divalent; endoribonucleases; bacteria; dna strand; peptide initiation factors; electrostatics; aquifex aeolicus |
| Journal Title: | Molecular Cell |
| Volume: | 19 |
| Issue: | 3 |
| ISSN: | 1097-2765 |
| Publisher: | Cell Press |
| Date Published: | 2005-08-05 |
| Start Page: | 405 |
| End Page: | 419 |
| Language: | English |
| DOI: | 10.1016/j.molcel.2005.07.011 |
| PUBMED: | 16061186 |
| PROVIDER: | scopus |
| PMCID: | PMC4689305 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 127" - "Export Date: 24 October 2012" - "CODEN: MOCEF" - "Source: Scopus" |
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