دورية أكاديمية
Key arginine residues in R2D2 dsRBD1 and dsRBD2 lead the siRNA recognition in Drosophila melanogaster RNAi pathway.
| العنوان: | Key arginine residues in R2D2 dsRBD1 and dsRBD2 lead the siRNA recognition in Drosophila melanogaster RNAi pathway. |
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| المؤلفون: | Aute R; CSIR - Centre for Cellular and Molecular Biology, Council of Scientific and Industrial Research, Uppal Road, Hyderabad 500007, Telangana, India; Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India., Waghela N; CSIR - Centre for Cellular and Molecular Biology, Council of Scientific and Industrial Research, Uppal Road, Hyderabad 500007, Telangana, India; Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India., Deshmukh MV; CSIR - Centre for Cellular and Molecular Biology, Council of Scientific and Industrial Research, Uppal Road, Hyderabad 500007, Telangana, India; Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India. Electronic address: mvdesh@ccmb.res.in. |
| المصدر: | Biophysical chemistry [Biophys Chem] 2024 Jul; Vol. 310, pp. 107247. Date of Electronic Publication: 2024 Apr 18. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Science B.V Country of Publication: Netherlands NLM ID: 0403171 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-4200 (Electronic) Linking ISSN: 03014622 NLM ISO Abbreviation: Biophys Chem Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Amsterdam : Elsevier Science B.V Original Publication: Amsterdam, North-Holland Pub. Co. |
| مواضيع طبية MeSH: | Drosophila melanogaster*/metabolism , Arginine*/chemistry , Arginine*/metabolism , Drosophila Proteins*/chemistry , Drosophila Proteins*/metabolism , Drosophila Proteins*/genetics , RNA, Small Interfering*/chemistry , RNA, Small Interfering*/metabolism , RNA Interference*, Animals ; RNA Helicases/metabolism ; RNA Helicases/chemistry ; RNA Helicases/genetics ; Protein Domains ; RNA-Binding Proteins |
| مستخلص: | In Drosophila melanogaster, Dcr-2:R2D2 heterodimer binds to the 21 nucleotide siRNA duplex to form the R2D2/Dcr-2 Initiator (RDI) complex, which is critical for the initiation of siRNA-induced silencing complex (RISC) assembly. During RDI complex formation, R2D2, a protein that contains three dsRNA binding domains (dsRBD), senses two aspects of the siRNA: thermodynamically more stable end (asymmetry sensing) and the 5'-phosphate (5'-P) recognition. Despite several detailed studies to date, the molecular determinants arising from R2D2 for performing these two tasks remain elusive. In this study, we have performed structural, biophysical, and biochemical characterization of R2D2 dsRBDs. We found that the solution NMR-derived structure of R2D2 dsRBD1 yielded a canonical α1-β1-β2-β3-α2 fold, wherein two arginine salt bridges provide additional stability to the R2D2 dsRBD1. Furthermore, we show that R2D2 dsRBD1 interacts with thermodynamically asymmetric siRNA duplex independent of its 5'-phosphorylation state, whereas R2D2 dsRBD2 prefers to interact with 5'-P siRNA duplex. The mutation of key arginine residues, R53 and R101, in concatenated dsRBDs of R2D2 results in a significant loss of siRNA duplex recognition. Our study deciphers the active roles of R2D2 dsRBDs by showing that dsRBD1 initiates siRNA recognition, whereas dsRBD2 senses 5'-phosphate as an authentic mark on functional siRNA. Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest. (Copyright © 2024 Elsevier B.V. All rights reserved.) |
| فهرسة مساهمة: | Keywords: Dcr-2; Drosophila melanogaster; R2D2; RNA interference (RNAi); siRNA |
| المشرفين على المادة: | 0 (R2D2 protein, Drosophila) |
| تواريخ الأحداث: | Date Created: 20240425 Date Completed: 20240521 Latest Revision: 20240521 |
| رمز التحديث: | 20240522 |
| DOI: | 10.1016/j.bpc.2024.107247 |
| PMID: | 38663122 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1873-4200 |
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| DOI: | 10.1016/j.bpc.2024.107247 |