Phosphate and R2D2 Restrict the Substrate Specificity of Dicer-2, an ATP-Driven Ribonuclease
Copyright policy )Phosphate and R2D2 Restrict the Substrate Specificity of Dicer-2, an ATP-Driven Ribonuclease
Drosophila Dicer-2 generates small interfering RNAs (siRNAs) from long double-stranded RNA (dsRNA), whereas Dicer-1 produces microRNAs (miRNAs) from pre-miRNA. What makes the two Dicers specific for their biological substrates? We find that purified Dicer-2 can efficiently cleave pre-miRNA, but that inorganic phosphate and the Dicer-2 partner protein R2D2 inhibit pre-miRNA cleavage. Dicer-2 contains C-terminal RNase III domains that mediate RNA cleavage and an N-terminal helicase motif, whose function is unclear. We show that Dicer-2 is a dsRNA-stimulated ATPase that hydrolyzes ATP to ADP; ATP hydrolysis is required for Dicer-2 to process long dsRNA, but not pre-miRNA. Wild-type Dicer-2, but not a mutant defective in ATP hydrolysis, can generate siRNAs faster than it can dissociate from a long dsRNA substrate. We propose that the Dicer-2 helicase domain uses ATP to generate many siRNAs from a single molecule of dsRNA before dissociating from its substrate.
- University of Massachusetts Medical School United States
- National Institute of Health Pakistan
- National Institutes of Health United States
- Research Triangle Park Foundation United States
- National Institute of Environmental Health Sciences United States
Ribonuclease III, Biophysics, Small Interfering, Biochemistry, Models, Biological, Dicer-2, Dicer-1, Phosphates, Substrate Specificity, Double-Stranded, Adenosine Triphosphate, Toxicology and Environmental Health, Animals, Drosophila Proteins, RNA Processing, Post-Transcriptional, RNA, Small Interfering, Molecular Biology, phosphate, RNA, Double-Stranded, Pharmacology, Hydrolysis, RNA-Binding Proteins, Cell Biology, processivity, R2D2, ATP, Adenosine Diphosphate, helicase, Kinetics, MicroRNAs, Drosophila melanogaster, and Structural Biology, RNA, RNA Helicases
Ribonuclease III, Biophysics, Small Interfering, Biochemistry, Models, Biological, Dicer-2, Dicer-1, Phosphates, Substrate Specificity, Double-Stranded, Adenosine Triphosphate, Toxicology and Environmental Health, Animals, Drosophila Proteins, RNA Processing, Post-Transcriptional, RNA, Small Interfering, Molecular Biology, phosphate, RNA, Double-Stranded, Pharmacology, Hydrolysis, RNA-Binding Proteins, Cell Biology, processivity, R2D2, ATP, Adenosine Diphosphate, helicase, Kinetics, MicroRNAs, Drosophila melanogaster, and Structural Biology, RNA, RNA Helicases
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