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紫外线对门戈病毒的影响:尿嘧啶二聚体的形成、衣壳的不稳定性和降解以及蛋白质与病毒RNA的共价连接。

Effect of ultraviolet light on mengovirus: formation of uracil dimers, instability and degradation of capsid, and covalent linkage of protein to viral RNA.

作者信息

Miller R L, Plagemann P G

出版信息

J Virol. 1974 Mar;13(3):729-39. doi: 10.1128/JVI.13.3.729-739.1974.

Abstract

UV irradiation of purified mengovirus resulted in a very rapid inactivation of the infectivity of the virions (D(37) [37% survival dose] = 700 ergs/mm(2)) which correlated in time with the formation of uracil dimers in the viral RNA. During the first 2 min of irradiation, an average of 1.7 uracil dimers were formed per PFU of virus inactivated. Hemagglutination activity of the virions began to decrease only after a lag period of about 5 min and at a much lower rate (D(37) = 84,000 ergs/mm(2)). This decrease coincided in time with the appearance of altered proteins in the capsid and a structural change in the capsid. Although 10- to 20-min irradiated virions appeared intact in the electron microscope and sedimented at 150S in sucrose density gradients, the RNA of the virions became accessible to RNase and extractable by low concentrations of sodium dodecyl sulfate, and the virions broke down upon equilibrium centrifugation in CsCl gradients. During longer periods of irradiation (30 to 60 min), a progressively greater proportion of the virions were converted to 14S protein particles and 80S ribonucleoprotein particles composed of intact viral RNA and about 30% of the capsid proteins, alpha, beta, and gamma. Empty capsids were not detectable at any time during 60 min of irradiation, by which time disruption of the virions was complete. Irradiation of complete virions also resulted in an increased sedimentation rate of the viral RNA and in the covalent linkage to the viral RNA of about 1% of the total capsid protein in the form of heterogeneous low-molecular-weight polypeptides. The two observations seem to be causally related, since irradiation of isolated viral RNA did not result in an increase in sedimentation rate of the RNA, even though uracil dimer formation in viral RNA occurred at about the same rate and to the same extent whether intact virions or viral RNA were irradiated.

摘要

纯化的 mengovirus 经紫外线照射后,病毒粒子的感染性迅速失活(D(37) [37%存活剂量] = 700 尔格/mm(2)),这与病毒 RNA 中尿嘧啶二聚体的形成在时间上相关。在照射的前 2 分钟内,每失活一个 PFU 的病毒平均形成 1.7 个尿嘧啶二聚体。病毒粒子的血凝活性仅在约 5 分钟的延迟期后开始下降,且下降速率低得多(D(37) = 84,000 尔格/mm(2))。这种下降在时间上与衣壳中蛋白质的改变以及衣壳结构的变化同时出现。尽管照射 10 至 20 分钟的病毒粒子在电子显微镜下看起来完整,且在蔗糖密度梯度中以 150S 沉降,但病毒粒子的 RNA 变得可被核糖核酸酶作用,并可用低浓度的十二烷基硫酸钠提取,并且在 CsCl 梯度中进行平衡离心时病毒粒子会分解。在更长时间的照射(30 至 60 分钟)期间,越来越大比例的病毒粒子转化为 14S 蛋白质颗粒和 80S 核糖核蛋白颗粒,后者由完整的病毒 RNA 和大约 30%的衣壳蛋白α、β和γ组成。在 60 分钟的照射期间,任何时候都检测不到空衣壳,此时病毒粒子的破坏已完成。完整病毒粒子的照射还导致病毒 RNA 的沉降速率增加,并且约 1%的总衣壳蛋白以异质低分子量多肽的形式与病毒 RNA 共价连接。这两个观察结果似乎存在因果关系,因为即使完整病毒粒子或病毒 RNA 被照射时病毒 RNA 中尿嘧啶二聚体的形成速率和程度大致相同,但分离的病毒 RNA 的照射并未导致 RNA 沉降速率增加。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b08/355360/33715fcb9992/jvirol00243-0188-a.jpg

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