Institute for Quantitative Biomedicine, Rutgers, The State University of New Jersey, Piscataway, NJ, USA.
Markey Center for Structural Biology, Department of Biological Sciences, Purdue University, West Lafayette, IN, USA.
Structure. 2020 Jun 2;28(6):604-612.e3. doi: 10.1016/j.str.2020.01.011. Epub 2020 Feb 10.
Members of the family Reoviridae package several copies of the viral polymerase complex into their capsid to carry out replication and transcription within viral particles. Classical single-particle reconstruction encounters difficulties resolving structures such as the intraparticle polymerase complex because refinement can converge to an incorrect map and because the map could depict a nonrepresentative subset of particles or an average of heterogeneous particles. Using the nine-segmented Fako virus, we tested hypotheses for the arrangement and number of polymerase complexes within the virion by measuring how well each hypothesis describes the set of cryoelectron microscopy images of individual viral particles. We find that the polymerase complex in Fako virus binds at ten possible sites despite having only nine genome segments. A single asymmetric configuration describes the arrangement of these complexes in both virions and genome-free capsids. Similarities between the arrangements of Reoviridae with 9, 10, and 11 segments indicate the generalizability of this architecture.
呼肠孤病毒科的成员将多个病毒聚合酶复合物包装到它们的衣壳中,以便在病毒颗粒内进行复制和转录。经典的单颗粒重建在解析聚合酶复合物等结构时会遇到困难,因为精修可能会收敛到错误的地图,并且地图可能描绘了非代表性的粒子子集或不均匀粒子的平均值。使用九段法科病毒,我们通过测量每个假设对单个病毒粒子的一组冷冻电镜图像的描述程度,来测试聚合酶复合物在病毒粒子内的排列和数量的假设。我们发现,尽管法科病毒只有九个基因组片段,但聚合酶复合物仍能结合在十个可能的位点上。一个单一的不对称构象描述了这些复合物在病毒粒子和无基因组衣壳中的排列。具有 9、10 和 11 个片段的呼肠孤病毒科的排列相似性表明了这种结构的普遍性。