The Making of a Slicer: Activation of Human Argonaute-1
Argonautes are the central protein component in small RNA silencing pathways. Of the four human Argonautes (hAgo1–hAgo4) only hAgo2 is an active slicer. We determined the structure of hAgo1 bound to endogenous copurified RNAs to 1.75 Å resolution and hAgo1 loaded with let-7 microRNA to 2.1 Å. Both s...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2013-06-01
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| Series: | Cell Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2211124713002660 |
| _version_ | 1818833257655959552 |
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| author | Christopher R. Faehnle Elad Elkayam Astrid D. Haase Gregory J. Hannon Leemor Joshua-Tor |
| author_facet | Christopher R. Faehnle Elad Elkayam Astrid D. Haase Gregory J. Hannon Leemor Joshua-Tor |
| author_sort | Christopher R. Faehnle |
| collection | DOAJ |
| description | Argonautes are the central protein component in small RNA silencing pathways. Of the four human Argonautes (hAgo1–hAgo4) only hAgo2 is an active slicer. We determined the structure of hAgo1 bound to endogenous copurified RNAs to 1.75 Å resolution and hAgo1 loaded with let-7 microRNA to 2.1 Å. Both structures are strikingly similar to the structures of hAgo2. A conserved catalytic tetrad within the PIWI domain of hAgo2 is required for its slicing activity. Completion of the tetrad, combined with a mutation on a loop adjacent to the active site of hAgo1, results in slicer activity that is substantially enhanced by swapping in the N domain of hAgo2. hAgo3, with an intact tetrad, becomes an active slicer by swapping the N domain of hAgo2 without additional mutations. Intriguingly, the elements that make Argonaute an active slicer involve a sophisticated interplay between the active site and more distant regions of the enzyme. |
| first_indexed | 2024-12-19T02:16:03Z |
| format | Article |
| id | doaj.art-3f6f678ec8e34787af29b6ea96cbef9e |
| institution | Directory Open Access Journal |
| issn | 2211-1247 |
| language | English |
| last_indexed | 2024-12-19T02:16:03Z |
| publishDate | 2013-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Cell Reports |
| spelling | doaj.art-3f6f678ec8e34787af29b6ea96cbef9e2022-12-21T20:40:24ZengElsevierCell Reports2211-12472013-06-01361901190910.1016/j.celrep.2013.05.033The Making of a Slicer: Activation of Human Argonaute-1Christopher R. Faehnle0Elad Elkayam1Astrid D. Haase2Gregory J. Hannon3Leemor Joshua-Tor4W.M. Keck Structural Biology Laboratory, Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USAW.M. Keck Structural Biology Laboratory, Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USAHoward Hughes Medical Institute, Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USAHoward Hughes Medical Institute, Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USAHoward Hughes Medical Institute, Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USAArgonautes are the central protein component in small RNA silencing pathways. Of the four human Argonautes (hAgo1–hAgo4) only hAgo2 is an active slicer. We determined the structure of hAgo1 bound to endogenous copurified RNAs to 1.75 Å resolution and hAgo1 loaded with let-7 microRNA to 2.1 Å. Both structures are strikingly similar to the structures of hAgo2. A conserved catalytic tetrad within the PIWI domain of hAgo2 is required for its slicing activity. Completion of the tetrad, combined with a mutation on a loop adjacent to the active site of hAgo1, results in slicer activity that is substantially enhanced by swapping in the N domain of hAgo2. hAgo3, with an intact tetrad, becomes an active slicer by swapping the N domain of hAgo2 without additional mutations. Intriguingly, the elements that make Argonaute an active slicer involve a sophisticated interplay between the active site and more distant regions of the enzyme.http://www.sciencedirect.com/science/article/pii/S2211124713002660 |
| spellingShingle | Christopher R. Faehnle Elad Elkayam Astrid D. Haase Gregory J. Hannon Leemor Joshua-Tor The Making of a Slicer: Activation of Human Argonaute-1 Cell Reports |
| title | The Making of a Slicer: Activation of Human Argonaute-1 |
| title_full | The Making of a Slicer: Activation of Human Argonaute-1 |
| title_fullStr | The Making of a Slicer: Activation of Human Argonaute-1 |
| title_full_unstemmed | The Making of a Slicer: Activation of Human Argonaute-1 |
| title_short | The Making of a Slicer: Activation of Human Argonaute-1 |
| title_sort | making of a slicer activation of human argonaute 1 |
| url | http://www.sciencedirect.com/science/article/pii/S2211124713002660 |
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