Raju R, Raju L, Kolakofsky D
Department of Microbiology, University of Geneva School of Medicine, Switzerland.
J Virol. 1989 Dec;63(12):5159-65. doi: 10.1128/JVI.63.12.5159-5165.1989.
The translational requirement to prevent premature termination during La Crosse virus S mRNA synthesis was found to be cell-type dependent. This requirement was present in the BHK, HEL, and Vero cell lines we examined, but not in C6/36 mosquito cells. The cell-dependent translational requirement could be reproduced in vitro by using either cell extracts or purified virions of BHK and C6/36 cells. In the BHK reactions, the polymerase terminated predominantly at nucleotide 175 in the absence of concurrent translation and required translation to read through this position. In the C6/36 reactions, however, the polymerase reads through nucleotide 175 efficiently independent of translation. Reconstitution studies suggested that the translational requirement was due to a factor(s) present in BHK, but not in C6/36, cells.
在拉克罗斯病毒S mRNA合成过程中,防止过早终止的翻译需求被发现是细胞类型依赖性的。我们检测的BHK、HEL和Vero细胞系中存在这种需求,但在C6/36蚊细胞中则不存在。通过使用BHK和C6/36细胞的细胞提取物或纯化病毒粒子,这种细胞依赖性翻译需求可以在体外重现。在BHK反应中,在没有同时进行翻译的情况下,聚合酶主要在核苷酸175处终止,并且需要翻译才能通读该位置。然而,在C6/36反应中,聚合酶能够高效地通读核苷酸175,而与翻译无关。重组研究表明,这种翻译需求是由于BHK细胞中存在而C6/36细胞中不存在的一种或多种因子所致。