Nonstructural N- and C-tails of Dbp2 confer the protein full helicase activities
| Autor: | Qin-Xia Song, Na-Nv Liu, Zhao-Xia Liu, Ying-Zi Zhang, Stephane Rety, Xi-Miao Hou, Xu-Guang Xi |
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| Přispěvatelé: | Northwest A and F University, Laboratoire de biologie et modélisation de la cellule (LBMC UMR 5239), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biotechnologie et Pharmacogénétique Appliquée (LBPA), École normale supérieure - Cachan (ENS Cachan)-Centre National de la Recherche Scientifique (CNRS), National Natural Science Foundation of China (32071291,32201042, 32071225, and 31870788)CNRS LIA ('Helicase-mediated G-quadruplex DNA unwinding and genome stability'). |
| Rok vydání: | 2023 |
| Předmět: |
disordered protein regions
RNA-protein interaction [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry Molecular Biology/Structural Biology [q-bio.BM] RNA helicase Dbp2 FRET Cell Biology [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biochemistry [q-bio.BM] RNA helicase Dbp2 disordered protein regions RNA-protein interaction X-ray crystallography FRET Molecular Biology Biochemistry X-ray crystallography |
| Zdroj: | Journal of Biological Chemistry Journal of Biological Chemistry, In press, ⟨10.1016/j.jbc.2023.104592⟩ |
| ISSN: | 0021-9258 1083-351X |
| DOI: | 10.1016/j.jbc.2023.104592 |
| Popis: | International audience; Human DDX5 and its yeast ortholog Dbp2 are ATP-dependent RNA helicases that play a keyrole in normal cell processes, cancer development and viral infection. The crystal structure ofthe RecA1-like domain of DDX5 is available, but the global structure of DDX5/Dbp2subfamily proteins remains to be elucidated. Here, we report the first X-ray crystal structuresof the Dbp2 helicase core alone and in complex with adenosine diphosphate nucleotide (ADP)at 3.22 Å and 3.05 Å resolutions, respectively. The structures of the ADP-bound post-hydrolysisstate and apo-state demonstrate the conformational changes that occur when the nucleotides arereleased. Our results showed that the helicase core of Dbp2 shifted between open and closedconformation in solution, but the unwinding activity was hindered when the helicase core wasrestricted to a single conformation. A small-angle X-ray scattering (SAXS) experiment showedthat the disordered amino- (N-) and carboxy- (C-) tails are flexible in solution. Truncationmutations confirmed that the N- and C-tails were critical for the nucleic acid binding, ATPase,and unwinding activities, with the C-tail being exclusively responsible for the annealing activity.Furthermore, we labeled the terminal tails to observe the conformational changes between thedisordered tails and the helicase core upon binding nucleic acid substrates. Specifically, wefound that the nonstructural N- and C-tails bind to RNA substrates and tether them to thehelicase core domain, thereby conferring full helicase activities to the Dbp2 protein. Thisdistinct structural characteristic provides new insight into the mechanism of DEAD-box RNAhelicases. |
| Databáze: | OpenAIRE |
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