| Title |
Structural determinants of miRNAs for RISC loading and slicer-independent unwinding
|
|---|---|
| Published in |
Nature Structural & Molecular Biology, August 2009
|
| DOI | 10.1038/nsmb.1630 |
| Pubmed ID | |
| Authors | |
| Abstract |
MicroRNAs (miRNAs) regulate expression of their target mRNAs through the RNA-induced silencing complex (RISC), which contains an Argonaute (Ago) family protein as a core component. In Drosophila melanogaster, miRNAs are generally sorted into Ago1-containing RISC (Ago1-RISC). We established a native gel system that can biochemically dissect the Ago1-RISC assembly pathway. We found that miRNA-miRNA* duplexes are loaded into Ago1 as double-stranded RNAs in an ATP-dependent fashion. In contrast, unexpectedly, unwinding of miRNA-miRNA* duplexes is a passive process that does not require ATP or slicer activity of Ago1. Central mismatches direct miRNA-miRNA* duplexes into pre-Ago1-RISC, whereas mismatches in the seed or guide strand positions 12-15 promote conversion of pre-Ago1-RISC into mature Ago1-RISC. Our findings show that unwinding of miRNAs is a precise mirror-image process of target recognition, and both processes reflect the unique geometry of RNAs in Ago proteins. |
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Mendeley demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 5 | 1% |
| United Kingdom | 4 | 1% |
| Luxembourg | 2 | <1% |
| Germany | 2 | <1% |
| Russia | 1 | <1% |
| Poland | 1 | <1% |
| Mexico | 1 | <1% |
| Japan | 1 | <1% |
| India | 1 | <1% |
| Other | 5 | 1% |
| Unknown | 352 | 94% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 107 | 29% |
| Researcher | 66 | 18% |
| Student > Master | 45 | 12% |
| Student > Bachelor | 26 | 7% |
| Student > Doctoral Student | 23 | 6% |
| Other | 57 | 15% |
| Unknown | 51 | 14% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 180 | 48% |
| Biochemistry, Genetics and Molecular Biology | 90 | 24% |
| Medicine and Dentistry | 12 | 3% |
| Chemistry | 10 | 3% |
| Neuroscience | 9 | 2% |
| Other | 18 | 5% |
| Unknown | 56 | 15% |