Fernandez-Larsson R, Patterson J L
Division of Infectious Diseases, Children's Hospital, Boston, Massachusetts.
J Gen Virol. 1989 Oct;70 ( Pt 10):2791-7. doi: 10.1099/0022-1317-70-10-2791.
We have studied the effect of phosphorylated ribavirin on the vesicular stomatitis virus (VSV) in vitro polymerase reaction by analysis of kinetic data obtained by varying the concentration of nucleoside triphosphates. The wild-type VSV had previously shown a competitive inhibition with the four natural nucleoside triphosphates with the use of ribavirin diphosphate (RDP) or ribavirin triphosphate (RTP). In contrast, when RDP (or RTP) was added to a transcription assay system using the polR1 mutant of VSV, a non-competitive or mixed type of inhibition was observed when the concentration of ATP was varied. Our results indicate that polR1 has an altered ATP function in addition to the previously described phenotypic characteristics of this mutant, which include synthesis of readthrough products of the leader/nucleocapsid (N) gene junction and a decreased ATP requirement for transcription. We have also studied CsCl-purified in vitro transcription products by primer-extending leader or N mRNA transcripts and found that the ratio of leader/N mRNA for VSV polR1 (1.3:1) was lower than values obtained previously for wild-type (3.7:1).
我们通过分析改变三磷酸核苷浓度所获得的动力学数据，研究了磷酸化利巴韦林对水疱性口炎病毒（VSV）体外聚合酶反应的影响。野生型VSV此前已显示，使用二磷酸利巴韦林（RDP）或三磷酸利巴韦林（RTP）时，会与四种天然三磷酸核苷产生竞争性抑制作用。相比之下，当将RDP（或RTP）添加到使用VSV的polR1突变体的转录测定系统中时，改变ATP浓度会观察到非竞争性或混合型抑制作用。我们的结果表明，除了该突变体先前描述的表型特征外，polR1还具有改变的ATP功能，这些特征包括前导/核衣壳（N）基因连接处通读产物的合成以及转录所需ATP的减少。我们还通过引物延伸前导或N mRNA转录本研究了氯化铯纯化的体外转录产物，发现VSV polR1的前导/N mRNA比例（1.3:1）低于先前野生型获得的值（3.7:1）。
