Academic Journal
Structural determinants of TAR RNA-DNA annealing in the absence and presence of HIV-1 nucleocapsid protein
| العنوان: | Structural determinants of TAR RNA-DNA annealing in the absence and presence of HIV-1 nucleocapsid protein |
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| المؤلفون: | Kanevsky, I., Chaminade, F., Chen, Y., Godet, J., Rene, B., Darlix, J.L., Mely, Y., Mauffret, O., Fosse, P. |
| المساهمون: | Laboratoire de Biophotonique et Pharmacologie - UMR 7213 (LBP), Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS) |
| المصدر: | ISSN: 0305-1048. |
| بيانات النشر: | HAL CCSD Oxford University Press |
| سنة النشر: | 2011 |
| المجموعة: | Archive ouverte HAL (Hyper Article en Ligne, CCSD - Centre pour la Communication Scientifique Directe) |
| مصطلحات موضوعية: | [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, [SDV.BC]Life Sciences [q-bio]/Cellular Biology |
| الوصف: | International audience ; Annealing of the TAR RNA hairpin to the cTAR DNA hairpin is required for the minus-strand transfer step of HIV-1 reverse transcription. HIV-1 nucleocapsid protein (NC) plays a crucial role by facilitating annealing of the complementary hairpins. To gain insight into the mechanism of NC-mediated TAR RNA-DNA annealing, we used structural probes (nucleases and potassium permanganate), gel retardation assays, fluorescence anisotropy and cTAR mutants under conditions allowing strand transfer. In the absence of NC, cTAR DNA-TAR RNA annealing depends on nucleation through the apical loops. We show that the annealing intermediate of the kissing pathway is a loop-loop kissing complex involving six base-pairs and that the apical stems are not destabilized by this loop-loop interaction. Our data support a dynamic structure of the cTAR hairpin in the absence of NC, involving equilibrium between both the closed conformation and the partially open 'Y' conformation. This study is the first to show that the apical and internal loops of cTAR are weak and strong binding sites for NC, respectively. NC slightly destabilizes the lower stem that is adjacent to the internal loop and shifts the equilibrium toward the 'Y' conformation exhibiting at least 12 unpaired nucleotides in its lower part. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| Relation: | hal-00667206; https://hal.science/hal-00667206; PUBMEDCENTRAL: PMC3185427 |
| DOI: | 10.1093/nar/gkr526 |
| الاتاحة: | https://hal.science/hal-00667206 https://doi.org/10.1093/nar/gkr526 |
| رقم الانضمام: | edsbas.EBDB2006 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1093/nar/gkr526 |
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