Benarroch Delphine, Egloff Marie-Pierre, Mulard Laurence, Guerreiro Catherine, Romette Jean-Louis, Canard Bruno
Centre National de la Recherche Scientifique and Universités d'Aix-Marseille I et II, UMR 6098, Architecture et Fonction des Macromolécules Biologiques, Ecole Supérieur d'Ingénieurs de Luminy-Case 925, 163 avenue de Luminy, 13288 Marseille cedex 9, France.
J Biol Chem. 2004 Aug 20;279(34):35638-43. doi: 10.1074/jbc.M400460200. Epub 2004 May 19.
Ribavirin is one of the few nucleoside analogues currently used in the clinic to treat RNA virus infections, but its mechanism of action remains poorly understood at the molecular level. Here, we show that ribavirin 5'-triphosphate inhibits the activity of the dengue virus 2'-O-methyltransferase NS5 domain (NS5MTase(DV)). Along with several other guanosine 5'-triphosphate analogues such as acyclovir, 5-ethynyl-1-beta-d-ribofuranosylimidazole-4-carboxamide (EICAR), and a series of ribose-modified ribavirin analogues, ribavirin 5'-triphosphate competes with GTP to bind to NS5MTase(DV). A structural view of the binding of ribavirin 5'-triphosphate to this enzyme was obtained by determining the crystal structure of a ternary complex consisting of NS5MTase(DV), ribavirin 5'-triphosphate, and S-adenosyl-l-homocysteine at a resolution of 2.6 A. These detailed atomic interactions provide the first structural insights into the inhibition of a viral enzyme by ribavirin 5'-triphosphate, as well as the basis for rational drug design of antiviral agents with improved specificity against the emerging flaviviruses.
利巴韦林是目前临床上用于治疗RNA病毒感染的少数核苷类似物之一，但其作用机制在分子水平上仍知之甚少。在此，我们表明利巴韦林5'-三磷酸抑制登革病毒2'-O-甲基转移酶NS5结构域（NS5MTase(DV)）的活性。与其他几种鸟苷5'-三磷酸类似物如阿昔洛韦、5-乙炔基-1-β-D-呋喃核糖基咪唑-4-甲酰胺（EICAR）以及一系列核糖修饰的利巴韦林类似物一起，利巴韦林5'-三磷酸与GTP竞争结合NS5MTase(DV)。通过确定由NS5MTase(DV)、利巴韦林5'-三磷酸和S-腺苷-L-高半胱氨酸组成的三元复合物的晶体结构，分辨率为2.6埃，获得了利巴韦林5'-三磷酸与该酶结合的结构视图。这些详细的原子相互作用首次提供了关于利巴韦林5'-三磷酸抑制病毒酶的结构见解，以及针对新兴黄病毒设计具有更高特异性的抗病毒药物的合理设计基础。
