論文 - 詳細
| RRC ID | 73755 |
|---|---|
| 著者 | Miyoshi K, Okada TN, Siomi H, Siomi MC. |
| タイトル | Characterization of the miRNA-RISC loading complex and miRNA-RISC formed in the Drosophila miRNA pathway. |
| ジャーナル | RNA |
| Abstract |
In Drosophila, miRNA is processed by Dicer-1 (DCR-1) from its precursor and loaded onto Argonaute1 (AGO1). AGO1 recognizes target mRNAs based on the miRNA sequence and suppresses the expression at post-transcriptional levels. GW182, a P-body component, localizes the AGO1 complex to processing bodies (P-bodies) where mRNA targets are decayed or stored. However, the details of the pathway remain elusive. In this study, two distinct types of AGO1-containing complexes from Drosophila Schneider2 (S2) cells were isolated and compared at the molecular level. The AGO1 complex with DCR-1 contained neither mature miRNA nor GW182 but exhibited pre-miRNA processing activity. The resultant mature RNA was loaded onto AGO1 within the complex. The AGO1 complex with GW182 excluded DCR-1, but possessed mature miRNA and showed no pre-miRNA processing activity. Thus, the AGO1-DCR-1 and AGO1-GW182 complexes correspond to miRLC (miRISC loading complex) and miRISC, respectively. The requirement for various domains of AGO1 in miRNA-loading and DCR-1/GW182 interaction was also examined. The Mid domain mutant (F2V2) interacted with DCR-1 but not with mature miRNA and GW182. The AGO1-PAZ mutant lacks the mature miRNA-binding ability but associates with either DCR-1 or GW182. The AGO1-PIWI mutant showed no Slicer activity but associates with mature miRNA. These results indicate that these domains are required differently for miRLC and miRISC formation in the miRNA pathway. |
| 巻・号 | 15(7) |
| ページ | 1282-91 |
| 公開日 | 2009-7-1 |
| DOI | 10.1261/rna.1541209 |
| PII | rna.1541209 |
| PMID | 19451544 |
| PMC | PMC2704077 |
| MeSH | Animals Argonaute Proteins Blotting, Northern Blotting, Western Drosophila Proteins / genetics Drosophila Proteins / metabolism Drosophila melanogaster / genetics* Drosophila melanogaster / growth & development Drosophila melanogaster / metabolism* Eukaryotic Initiation Factors Fluorescent Antibody Technique Immunoprecipitation MicroRNAs / genetics MicroRNAs / metabolism* RNA Helicases / genetics RNA Helicases / metabolism RNA, Messenger / metabolism* RNA, Small Interfering / pharmacology RNA-Induced Silencing Complex / genetics RNA-Induced Silencing Complex / metabolism* Ribonuclease III / genetics Ribonuclease III / metabolism Signal Transduction* |
| IF | 4.32 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 各媒体での言及数の合計 | 0 |
| リソース情報 | |
| ショウジョウバエ | 10004Ab-1 10005Ab-1 10014Ab-1 |