PloS one, 2012-12, Vol.7 (12), p.e51829-e51829
2012
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- Autor(en) / Beteiligte
- Titel
- The role of human Dicer-dsRBD in processing small regulatory RNAs
- Ist Teil von
- PloS one, 2012-12, Vol.7 (12), p.e51829-e51829
- Ort / Verlag
- United States: Public Library of Science
- Erscheinungsjahr
- 2012
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- One of the most exciting recent developments in RNA biology has been the discovery of small non-coding RNAs that affect gene expression through the RNA interference (RNAi) mechanism. Two major classes of RNAs involved in RNAi are small interfering RNA (siRNA) and microRNA (miRNA). Dicer, an RNase III enzyme, plays a central role in the RNAi pathway by cleaving precursors of both of these classes of RNAs to form mature siRNAs and miRNAs, which are then loaded into the RNA-induced silencing complex (RISC). miRNA and siRNA precursors are quite structurally distinct; miRNA precursors are short, imperfect hairpins while siRNA precursors are long, perfect duplexes. Nonetheless, Dicer is able to process both. Dicer, like the majority of RNase III enzymes, contains a dsRNA binding domain (dsRBD), but the data are sparse on the exact role this domain plays in the mechanism of Dicer binding and cleavage. To further explore the role of human Dicer-dsRBD in the RNAi pathway, we determined its binding affinity to various RNAs modeling both miRNA and siRNA precursors. Our study shows that Dicer-dsRBD is an avid binder of dsRNA, but its binding is only minimally influenced by a single-stranded - double-stranded junction caused by large terminal loops observed in miRNA precursors. Thus, the Dicer-dsRBD contributes directly to substrate binding but not to the mechanism of differentiating between pre-miRNA and pre-siRNA. In addition, NMR spin relaxation and MD simulations provide an overview of the role that dynamics contribute to the binding mechanism. We compare this current study with our previous studies of the dsRBDs from Drosha and DGCR8 to give a dynamic profile of dsRBDs in their apo-state and a mechanistic view of dsRNA binding by dsRBDs in general.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1932-6203eISSN: 1932-6203DOI: 10.1371/journal.pone.0051829Titel-ID: cdi_plos_journals_1327136436
- Format
- –
- Schlagworte
- Amino Acid Sequence, Base Sequence, Binding, Biology, Computer simulation, Crystal structure, Double-stranded RNA, Drosophila, Enzymes, Gene expression, Genes, Giardia, Humans, Hydrolases, Insects, Kinases, MicroRNA, miRNA, Models, Molecular, Molecular biology, Molecular Sequence Data, NMR, Nuclear magnetic resonance, Nucleic Acid Conformation, Protein Binding, Protein Conformation, Protein Interaction Domains and Motifs, Ribonuclease, Ribonuclease III, Ribonuclease III - chemistry, Ribonuclease III - metabolism, Ribonucleic acid, RNA, RNA Precursors - chemistry, RNA Precursors - genetics, RNA Precursors - metabolism, RNA, Double-Stranded - genetics, RNA, Double-Stranded - metabolism, RNA, Small Interfering - chemistry, RNA, Small Interfering - genetics, RNA, Small Interfering - metabolism, RNA-induced silencing complex, RNA-mediated interference, Sedimentation & deposition, siRNA, Substrates