Structure of yeast Argonaute with guide RNA Journal Article
| Authors: | Nakanishi, K.; Weinberg, D. E.; Bartel, D. P.; Patel, D. J. |
| Article Title: | Structure of yeast Argonaute with guide RNA |
| Abstract: | The RNA-induced silencing complex, comprising Argonaute and guide RNA, mediates RNA interference. Here we report the 3.2A° crystal structure of Kluyveromyces polysporus Argonaute (KpAGO) fortuitously complexed with guide RNA originating from small-RNA duplexes autonomously loaded by recombinant KpAGO. Despite their diverse sequences, guide-RNA nucleotides 1-8 are positioned similarly, with sequence-independent contacts to bases, phosphates and 2-2-hydroxyl groups pre-organizing the backbone of nucleotides 2-8 in a near-A-form conformation. Compared with prokaryotic Argonautes, KpAGO has numerous surface-exposed insertion segments, with a cluster of conserved insertions repositioning the N domain to enable full propagation of guide-target pairing. Compared with Argonautes in inactive conformations, KpAGO has a hydrogen-bond network that stabilizes an expanded and repositioned loop, which inserts an invariant glutamate into the catalytic pocket. Mutation analyses and analogies to ribonuclease H indicate that insertion of this glutamate finger completes a universally conserved catalytic tetrad, thereby activating Argonaute for RNA cleavage. © 2012 Macmillan Publishers Limited. All rights reserved. |
| Keywords: | sequence analysis; mutation; nonhuman; protein conformation; protein domain; phosphatase; rna interference; enzyme activity; rna; molecular sequence data; amino terminal sequence; nucleotide sequence; base sequence; crystal structure; hydrogen bond; models, molecular; crystallography, x-ray; gene silencing; yeast; saccharomycetales; protein structure; gene insertion; glutamic acid; catalytic domain; argonaute protein; phosphate; molecular conformation; kluyveromyces; rna cleavage; biocatalysis; nucleotide; ribonuclease h; fungal proteins; hydroxyl group; eukaryotic cells; argonaute proteins; guide rna; rna, guide |
| Journal Title: | Nature |
| Volume: | 486 |
| Issue: | 7403 |
| ISSN: | 0028-0836 |
| Publisher: | Nature Publishing Group |
| Date Published: | 2012-06-20 |
| Start Page: | 368 |
| End Page: | 374 |
| Language: | English |
| DOI: | 10.1038/nature11211 |
| PROVIDER: | scopus |
| PUBMED: | 22722195 |
| PMCID: | PMC3853139 |
| DOI/URL: | |
| Notes: | --- - "Export Date: 2 July 2012" - "CODEN: NATUA" - "Source: Scopus" |
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