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The RNA-binding proteins HYL1 and SE promote accurate in vitro processing of pri-miRNA by DCL1
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| Zeitschriftentitel: | Proceedings of the National Academy of Sciences |
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| Personen und Körperschaften: | , , |
| In: | Proceedings of the National Academy of Sciences, 105, 2008, 29, S. 9970-9975 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Proceedings of the National Academy of Sciences
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| Schlagwörter: |
| Zusammenfassung: | <jats:p> The results of genetic studies in <jats:italic>Arabidopsis</jats:italic> indicate that three proteins, the RNase III DICER-Like1 (DCL1), the dsRNA-binding protein HYPONASTIC LEAVES1 (HYL1), and the C2H2 Zn-finger protein SERRATE (SE), are required for the accurate processing of microRNA (miRNA) precursors in the plant cell nucleus. To elucidate the biochemical mechanism of miRNA processing, we developed an <jats:italic>in vitro</jats:italic> miRNA processing assay using purified recombinant proteins. We find that DCL1 alone releases 21-nt short RNAs from dsRNA as well as synthetic miR167b precursor RNAs. However, correctly processed miRNAs constitute a minority of the cleavage products. We show that recombinant HYL1 and SE proteins accelerate the rate of DCL1-mediated cleavage of pre- and pri-miR167b substrates and promote accurate processing. </jats:p> |
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| Umfang: | 9970-9975 |
| ISSN: |
0027-8424
1091-6490 |
| DOI: | 10.1073/pnas.0803356105 |