CNRS, AFMB UMR 7257, 13288, Marseille, France; Aix-Marseille Université, AFMB UMR 7257, 13288, Marseille, France.
Center for Genome Sciences, US Army Medical Research Institute of Infectious Disease, 1425 Porter Street, Fort Detrick, MD 21702, United States.
Virus Res. 2015 Apr 2;201:94-100. doi: 10.1016/j.virusres.2015.02.018. Epub 2015 Feb 25.
Random transposon insertions in viral genomes can be used to reveal genomic regions important for virus replication. We used these genomic data to evaluate at the protein level the effect of such insertions on the Venezuelan Equine Encephalitis Virus nsP3 macro domain. The structural analysis showed that transposon insertions occur mainly in loops connecting the secondary structure elements. Some of the insertions leading to a temperature sensitive viral phenotype (ts) are close to the cleavage site between nsP2 and nsP3 or the ADP-ribose binding site, two important functions of the macro domain. Using four mutants mimicking the transposon insertions, we confirmed that these insertions can affect the macro domain properties without disrupting the overall structure of the protein.
病毒基因组中的随机转座子插入可用于揭示对病毒复制重要的基因组区域。我们使用这些基因组数据来评估委内瑞拉马脑炎病毒 nsP3 大结构域中这种插入对蛋白质水平的影响。结构分析表明，转座子插入主要发生在连接二级结构元件的环中。一些导致温度敏感病毒表型 (ts) 的插入靠近 nsP2 和 nsP3 之间的切割位点或 ADP-核糖结合位点，这是大结构域的两个重要功能。使用四个模拟转座子插入的突变体，我们证实这些插入可以影响大结构域的特性，而不会破坏蛋白质的整体结构。
