Potisopon Supanee, Ferron François, Fattorini Véronique, Selisko Barbara, Canard Bruno
Aix-Marseille Université, AFMB (Laboratoire d'Architecture et Fonction de Macromolécules Biologiques) UMR 7257, 163 Avenue de Luminy, 13288 Marseille, France; CNRS, AFMB UMR 7257, 163 Avenue de Luminy, 13288 Marseille, France.
Aix-Marseille Université, AFMB (Laboratoire d'Architecture et Fonction de Macromolécules Biologiques) UMR 7257, 163 Avenue de Luminy, 13288 Marseille, France; CNRS, AFMB UMR 7257, 163 Avenue de Luminy, 13288 Marseille, France.
Antiviral Res. 2017 Apr;140:25-36. doi: 10.1016/j.antiviral.2016.12.021. Epub 2016 Dec 30.
In targeting the essential viral RNA-dependent RNA-polymerase (RdRp), nucleotide analogues play a major role in antiviral therapies. In the Flaviviridae family, the hepatitis C virus (HCV) can be eradicated from chronically infected patients using a combination of drugs which generally include the 2'-modified uridine analogue Sofosbuvir, delivered as nucleotide prodrug. Dengue and Zika viruses are emerging flaviviruses whose RdRp is closely related to that of HCV, yet no nucleoside drug has been clinically approved for these acute infections. We have purified dengue and Zika virus full-length NS5, the viral RdRps, and used them to assemble a stable binary complex made of NS5 and virus-specific RNA primer/templates. The complex was used to assess the selectivity of NS5 towards nucleotide analogues bearing modifications at the 2'-position. We show that dengue and Zika virus RdRps exhibit the same discrimination pattern: 2'-O-Me > 2'-C-Me-2'-F > 2'-C-Me nucleoside analogues, unlike HCV RdRp for which the presence of the 2'-F is beneficial rendering the discrimination pattern 2'-O-Me > 2'-C-Me ≥ 2'-C-Me-2'-F. Both 2'-C-Me and 2'-C-Me-2'-F analogues act as non-obligate RNA chain terminators. The dengue and Zika NS5 nucleotide selectivity towards 2'-modified NTPs mirrors potency of the corresponding analogues in infected cell cultures.
在靶向关键的病毒RNA依赖性RNA聚合酶（RdRp）方面，核苷酸类似物在抗病毒治疗中发挥着重要作用。在黄病毒科中，丙型肝炎病毒（HCV）可通过使用通常包括2'-修饰的尿苷类似物索磷布韦（作为核苷酸前药给药）的联合药物，从慢性感染患者体内根除。登革热病毒和寨卡病毒是新兴的黄病毒，它们的RdRp与HCV的RdRp密切相关，但尚无核苷类药物被临床批准用于这些急性感染。我们已经纯化了登革热病毒和寨卡病毒的全长NS5（病毒RdRp），并利用它们组装了由NS5和病毒特异性RNA引物/模板组成的稳定二元复合物。该复合物用于评估NS5对在2'-位带有修饰的核苷酸类似物的选择性。我们发现，登革热病毒和寨卡病毒的RdRp表现出相同的区分模式：2'-O-甲基 > 2'-C-甲基-2'-氟 > 2'-C-甲基核苷类似物，这与HCV RdRp不同，对于HCV RdRp而言，2'-氟的存在是有益的，其区分模式为2'-O-甲基 > 2'-C-甲基 ≥ 2'-C-甲基-2'-氟。2'-C-甲基和2'-C-甲基-2'-氟类似物均作为非必需的RNA链终止剂。登革热病毒和寨卡病毒NS5对2'-修饰的NTP的核苷酸选择性反映了相应类似物在感染细胞培养物中的效力。
