Ribavirin causes error catastrophe during Hantaan virus replication
Ribavirin causes error catastrophe during Hantaan virus replication
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DOI：
10.1128/jvi.77.1.481-488.2003
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发表时间：
2003-01-01
影响因子：
5.4
通讯作者：
Jonsson, CB
中科院分区：
文献类型：
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作者：
Severson, WE;Schmaljohn, CS;Jonsson, CB
除利巴韦林外，尚未发现其他治疗汉他病毒病的抗病毒药物。众所周知，利巴韦林将抑制传染性汉滩病毒（HTNV）的产生;然而，其作用机制尚不清楚。为了表征利巴韦林对HTNV的抑制作用，在HTNV感染的Vero E6细胞处理后3天测量病毒RNA、蛋白质和感染性颗粒的水平。病毒唑处理的HTNV感染细胞在感染后第一天cRNA、病毒RNA和mRNA的水平略有下降。在第2天，cRNA和病毒RNA的量增加至未处理的HTNV感染细胞所观察到的量，而mRNA水平在第2天和第3天更大程度地降低。尽管发现的S-段mRNA，虽然低，三个病毒蛋白-核衣壳（N）蛋白和糖蛋白G1和G2-不能检测到利巴韦林处理的细胞免疫组化。为了检验这些效应是由利巴韦林掺入新生RNA引起的并且由此产生的“错误灾难”正在发生的假设，我们从第3天起对利巴韦林处理或未处理的细胞的S-片段cRNA/mRNA进行克隆和测序。我们发现在利巴韦林存在下合成的病毒RNA中突变频率很高（9.5/1，000个核苷酸）。因此，在药物存在下产生的转录物没有功能。这些结果表明，利巴韦林的作用机制在于挑战汉坦病毒聚合酶的保真度，从而导致错误灾难。
Except for ribavirin, no other antiviral drugs for treating hantaviral diseases have been identified. It is well established that ribavirin will inhibit the production of infectious Hantaan virus (HTNV); however, its mechanism of action is unknown. To characterize the inhibitory effect of ribavirin on HTNV, the levels of viral RNAs, proteins, and infectious particles were measured for 3 days posttreatment of HTNV-infected Vero E6 cells. HTNV-infected cells treated with ribavirin showed a slight reduction in the levels of cRNA, viral RNA, and mRNA populations on the first day postinfection. The amount of cRNA and viral RNA increased to that observed for untreated HTNV-infected cells on day 2, whereas mRNA levels were more greatly reduced on days 2 and 3. Despite the finding of S-segment mRNA, albeit low, three of the viral proteins-nucleocapsid (N) protein and glycoproteins G1 and G2-could not be detected by immunohistochemistry in ribavirin-treated cells. To test the hypothesis that these effects were caused by incorporation of ribavirin into nascent RNA and a resultant "error catastrophe" was occurring, we cloned and sequenced the S-segment cRNA/mRNA from ribavirin-treated or untreated cells from day 3. We found a high mutation frequency (9.5/1,000 nucleotides) in viral RNA synthesized in the presence of ribavirin. Hence, the transcripts produced in the presence of the drug were not functional. These results suggest that ribavirin's mechanism of action lies in challenging the fidelity of the hantavirus polymerase, which causes error catastrophe.