Willkomm, Sarah and Jakob, Leonhard and Kramm, Kevin and Graus, Veronika and Neumeier, Julia and Meister, Gunter and Grohmann, Dina (2022) Single-molecule FRET uncovers hidden conformations and dynamics of human Argonaute 2. NATURE COMMUNICATIONS, 13 (1): 3825. ISSN , 2041-1723
Full text not available from this repository. (Request a copy)Abstract
Human Argonaute 2 (hAgo2) constitutes the functional core of the RNA interference pathway. Guide RNAs direct hAgo2 to target mRNAs, which ultimately leads to hAgo2-mediated mRNA degradation or translational inhibition. Here, we combine site-specifically labeled hAgo2 with time-resolved single-molecule FRET measurements to monitor conformational states and dynamics of hAgo2 and hAgo2-RNA complexes in solution that remained elusive so far. We observe dynamic anchoring and release of the guide's 3'-end from the PAZ domain during the stepwise target loading process even with a fully complementary target. We find differences in structure and dynamic behavior between partially and fully paired canonical hAgo2-guide/target complexes and the miRNA processing complex formed by hAgo2 and pre-miRNA451. Furthermore, we detect a hitherto unknown conformation of hAgo2-guide/target complexes that poises them for target-directed miRNA degradation. Taken together, our results show how the conformational flexibility of hAgo2-RNA complexes determines function and the fate of the ribonucleoprotein particle. Single-molecule FRET measurements provide detailed insights into the conformational states and dynamics of human Argonaute 2 that are required for its function at the core of the eukaryotic RNA silencing pathway.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | STRUCTURAL BASIS; GUIDE RNA; CRYSTAL-STRUCTURE; TARGET RNAS; MICRORNA; MESSENGER; PROTEINS; RISC; RECOGNITION; CLEAVAGE; |
| Subjects: | 500 Science > 570 Life sciences 600 Technology > 615 Pharmacy |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie I > Prof. Dr. Gunter Meister Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Mikrobiologie (Archaeenzentrum) > Prof. Dr. Dina Grohmann Chemistry and Pharmacy > Institute of Pharmacy > Pharmacology and Toxicology (Prof. Schlossmann, formerly Prof. Seifert) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 10 Oct 2023 14:29 |
| Last Modified: | 10 Oct 2023 14:29 |
| URI: | https://pred.uni-regensburg.de/id/eprint/56459 |
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