The ambiguous base-pairing and high substrate efficiency of T-705 (Favipiravir) Ribofuranosyl 5'-triphosphate towards influenza A virus polymerase.
The ambiguous base-pairing and high substrate efficiency of T-705 (Favipiravir) Ribofuranosyl 5'-triphosphate towards influenza A virus polymerase.
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T-705（法匹拉韦）呋喃核糖基 5'-三磷酸对甲型流感病毒聚合酶的模糊碱基配对和高底物效率。
DOI：
10.1371/journal.pone.0068347
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Deval J
中科院分区：
文献类型：
--
作者：
Jin Z;Smith LK;Rajwanshi VK;Kim B;Deval J
T-705（法匹拉韦）是一种广谱抗病毒分子，目前处于临床开发后期，用于治疗流感病毒感染。尽管认为T-705的效力是由其可能具有致突变性的呋喃核糖基三磷酸（T-705 RTP）代谢产物介导的，但尚未阐明与甲型流感病毒聚合酶（IAVpol）的确切分子相互作用。在这里，我们开发了一种生物化学测定来测量IAVpol在延伸模式中的核苷酸掺入动力学。在该测定中，T-705 RTP被IAVpol识别为作为鸟苷和腺苷类似物掺入RNA的有效底物。与天然GTP和ATP相比，T-705 RTP的区分度分别约为19倍和30倍。尽管单次掺入核糖核苷酸单磷酸形式的T-705不能有效地阻断RNA合成，但两次连续掺入事件阻止了进一步的引物延伸。相比之下，3′-脱氧GTP导致链立即终止，但被酶掺入的效率较低，相对于天然GTP的区分度为4，900倍。总的来说，这些结果提供了第一个详细的生物化学表征，以评估底物效率和核苷酸类似物对流感病毒聚合酶的抑制效力。T-705 RTP的不明确碱基配对与低区分度的组合为T-705对流感病毒的体外诱变作用提供了机制基础。
T-705 (Favipiravir) is a broad-spectrum antiviral molecule currently in late stage clinical development for the treatment of influenza virus infection. Although it is believed that T-705 potency is mediated by its ribofuranosyl triphosphate (T-705 RTP) metabolite that could be mutagenic, the exact molecular interaction with the polymerase of influenza A virus (IAVpol) has not been elucidated. Here, we developed a biochemical assay to measure the kinetics of nucleotide incorporation by IAVpol in the elongation mode. In this assay, T-705 RTP was recognized by IAVpol as an efficient substrate for incorporation to the RNA both as a guanosine and an adenosine analog. Compared to natural GTP and ATP, the discrimination of T-705 RTP was about 19- and 30-fold, respectively. Although the single incorporation of the ribonucleotide monophosphate form of T-705 did not efficiently block RNA synthesis, two consecutive incorporation events prevented further primer extension. In comparison, 3′-deoxy GTP caused immediate chain termination but was incorporated less efficiently by the enzyme, with a discrimination of 4,900-fold relative to natural GTP. Collectively, these results provide the first detailed biochemical characterization to evaluate the substrate efficiency and the inhibition potency of nucleotide analogs against influenza virus polymerase. The combination of ambiguous base-pairing with low discrimination of T-705 RTP provides a mechanistic basis for the in vitro mutagenic effect of T-705 towards influenza virus.
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影响因子：
3.7
作者：
Gowen BB;Smee DF;Wong MH;Hall JO;Jung KH;Bailey KW;Stevens JR;Furuta Y;Morrey JD
通讯作者：
Morrey JD
影响因子：
5.4
作者：
Hwang, JS;Yamada, K;Ishihama, A
通讯作者：
Ishihama, A
影响因子：
2.9
作者：
Arnold, JJ;Cameron, CE
通讯作者：
Cameron, CE
DOI：
10.1073/pnas.84.22.8140
发表时间：
1987-11-01
影响因子：
11.1
作者：
HSU, MT;PARVIN, JD;PALESE, P
通讯作者：
PALESE, P
影响因子：
3.7
作者：
Aggarwal S;Bradel-Tretheway B;Takimoto T;Dewhurst S;Kim B
通讯作者：
Kim B