Ciaccafava Alexandre, Lartigue Audrey, Mansuelle Pascal, Jeudy Sandra, Abergel Chantal
Information Génomique and Structurale, CNRS UPR 2589, Institut de Microbiologie de la Méditerranée, Aix-Marseille Université, Marseille, France.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Aug 1;67(Pt 8):922-5. doi: 10.1107/S1744309111020847. Epub 2011 Jul 20.
Mimivirus is the prototype of a new family (the Mimiviridae) of nucleocytoplasmic large DNA viruses (NCLDVs), which already include the Poxviridae, Iridoviridae, Phycodnaviridae and Asfarviridae. Mimivirus specifically replicates in cells from the genus Acanthamoeba. Proteomic analysis of purified mimivirus particles revealed the presence of many subunits of the DNA-directed RNA polymerase II complex. A fully functional pre-transcriptional complex appears to be loaded in the virions, allowing mimivirus to initiate transcription within the host cytoplasm immediately upon infection independently of the host nuclear apparatus. To fully understand this process, a systematic study of mimivirus proteins that are predicted (by bioinformatics) or suspected (by proteomic analysis) to be involved in transcription was initiated by cloning and expressing them in Escherichia coli in order to determine their three-dimensional structures. Here, preliminary crystallographic analysis of the recombinant L544 protein is reported. The crystals belonged to the orthorhombic space group C222(1) with one monomer per asymmetric unit. A MAD data set was used for preliminary phasing using the selenium signal present in a selenomethionine-substituted protein crystal.
巨型病毒是核质大DNA病毒(NCLDVs)新家族(巨型病毒科)的原型,该家族已包括痘病毒科、虹彩病毒科、藻DNA病毒科和非洲猪瘟病毒科。巨型病毒专门在棘阿米巴属的细胞中复制。对纯化的巨型病毒颗粒进行蛋白质组分析,发现了DNA指导的RNA聚合酶II复合物的许多亚基。一个功能齐全的转录前复合物似乎被装载到病毒粒子中,使巨型病毒在感染后能够立即在宿主细胞质内启动转录,而无需宿主核装置的参与。为了全面了解这一过程,通过在大肠杆菌中克隆和表达预测(通过生物信息学)或怀疑(通过蛋白质组分析)参与转录的巨型病毒蛋白,启动了一项系统研究,以确定它们的三维结构。在此,报告了重组L544蛋白的初步晶体学分析。晶体属于正交空间群C222(1),每个不对称单元有一个单体。使用硒代甲硫氨酸取代的蛋白质晶体中存在的硒信号,利用MAD数据集进行初步相位分析。