Structure of yeast Argonaute with guide RNA
The RNA-induced silencing complex, comprising Argonaute and guide RNA, mediates RNA interference. Here we report the 3.2 Å crystal structure of Kluyveromyces polysporus Argonaute (KpAGO) fortuitously complexed with guide RNA originating from small-RNA duplexes autonomously loaded and processed by re...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | en_US |
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Nature Publishing Group
2013
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| Online Access: | http://hdl.handle.net/1721.1/83083 https://orcid.org/0000-0002-3872-2856 |
| _version_ | 1811080502094856192 |
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| author | Nakanishi, Kotaro Weinberg, David E. Bartel, David Patel, Dinshaw J. |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Nakanishi, Kotaro Weinberg, David E. Bartel, David Patel, Dinshaw J. |
| author_sort | Nakanishi, Kotaro |
| collection | MIT |
| description | The RNA-induced silencing complex, comprising Argonaute and guide RNA, mediates RNA interference. Here we report the 3.2 Å crystal structure of Kluyveromyces polysporus Argonaute (KpAGO) fortuitously complexed with guide RNA originating from small-RNA duplexes autonomously loaded and processed by recombinant KpAGO. Despite their diverse sequences, guide-RNA nucleotides 1–8 are positioned similarly, with sequence-independent contacts to bases, phosphates and 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. |
| first_indexed | 2024-09-23T11:32:40Z |
| format | Article |
| id | mit-1721.1/83083 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T11:32:40Z |
| publishDate | 2013 |
| publisher | Nature Publishing Group |
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| spelling | mit-1721.1/830832022-09-27T20:13:29Z Structure of yeast Argonaute with guide RNA Nakanishi, Kotaro Weinberg, David E. Bartel, David Patel, Dinshaw J. Massachusetts Institute of Technology. Department of Biology Whitehead Institute for Biomedical Research Weinberg, David E. Bartel, David The RNA-induced silencing complex, comprising Argonaute and guide RNA, mediates RNA interference. Here we report the 3.2 Å crystal structure of Kluyveromyces polysporus Argonaute (KpAGO) fortuitously complexed with guide RNA originating from small-RNA duplexes autonomously loaded and processed by recombinant KpAGO. Despite their diverse sequences, guide-RNA nucleotides 1–8 are positioned similarly, with sequence-independent contacts to bases, phosphates and 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. National Institutes of Health (U.S.) (grant AI068776) National Institutes of Health (U.S.) (grant GM61835) Human Frontier Science Program (Strasbourg, France) (Long-term Fellowship) Japan Society for the Promotion of Science Howard Hughes Medical Institute (Investigator) National Science Foundation (U.S.) (Graduate Research Fellowship) 2013-12-19T19:43:50Z 2013-12-19T19:43:50Z 2012-06 2012-04 Article http://purl.org/eprint/type/JournalArticle 0028-0836 1476-4687 http://hdl.handle.net/1721.1/83083 Nakanishi, Kotaro, David E. Weinberg, David P. Bartel, and Dinshaw J. Patel. “Structure of yeast Argonaute with guide RNA.” Nature 486, no. 7403 (June 20, 2012): 368-374. https://orcid.org/0000-0002-3872-2856 en_US http://dx.doi.org/10.1038/nature11211 Nature Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf Nature Publishing Group PMC |
| spellingShingle | Nakanishi, Kotaro Weinberg, David E. Bartel, David Patel, Dinshaw J. Structure of yeast Argonaute with guide RNA |
| title | Structure of yeast Argonaute with guide RNA |
| title_full | Structure of yeast Argonaute with guide RNA |
| title_fullStr | Structure of yeast Argonaute with guide RNA |
| title_full_unstemmed | Structure of yeast Argonaute with guide RNA |
| title_short | Structure of yeast Argonaute with guide RNA |
| title_sort | structure of yeast argonaute with guide rna |
| url | http://hdl.handle.net/1721.1/83083 https://orcid.org/0000-0002-3872-2856 |
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