LACK OF EVIDENCE FOR PROOFREADING MECHANISMS ASSOCIATED WITH AN RNA VIRUS POLYMERASE
LACK OF EVIDENCE FOR PROOFREADING MECHANISMS ASSOCIATED WITH AN RNA VIRUS POLYMERASE
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DOI：
10.1016/0378-1119(92)90216-c
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发表时间：
1992-12-15
期刊：
影响因子：
3.5
通讯作者：
HOLLAND, JJ
中科院分区：
文献类型：
--
作者：
STEINHAUER, DA;DOMINGO, E;HOLLAND, JJ
对水疱性口炎病毒病毒粒子相关RNA聚合酶的体外保真度进行了定量分析，以确定单个保守的病毒RNA位点和通常高的体外碱基错误掺入错误频率(约为1。在该（鸟嘌呤）位点观察到10（-3）只。我们通过改变下一个核苷三磷酸的浓度、加入四种天然RNA核苷的核苷[1-硫代]三磷酸类似物以及在体外聚合酶反应中改变焦磷酸盐的浓度来寻找RNA 3’—bbb50’核酸外切酶校对机制的证据。这些干扰都没有对研究位点的病毒RNA聚合酶保真度产生很大影响。这些结果不能证明校对与RNA病毒的病毒粒子复制酶相关的外切酶活性。他们认为RNA病毒的复制通常是容易出错的，因为RNA复制酶碱基错配的校对效率非常低，或者根本没有。
The in vitro fidelity of the virion-associated RNA polymerase of vesicular stomatitis virus was quantitated for a single conserved viral RNA site and the usual high in vitro base misincorporation error frequencies (approx. 10(-3)) were observed at this (guanine) site. We sought evidence for RNA 3'-->5' exonuclease proofreading mechanisms by varying the concentrations of the next nucleoside triphosphate, by incorporation of nucleoside[ 1-thio]triphosphate analogues of the four natural RNA nucleosides, and by varying the concentrations of pyrophosphate in the in vitro polymerase reaction. None of these perturbations greatly affected viral RNA polymerase fidelity at the site studied. These results fail to show evidence for proofreading exonuclease activity associated with the virion replicase of an RNA virus. They suggest that RNA virus replication might generally be error-prone, because RNA replicase base misincorporations are proofread very inefficiently or not at all.