Khan Mirzaei Mohammadali, Eriksson Harald, Kasuga Kie, Haggård-Ljungquist Elisabeth, Nilsson Anders S
Department of Molecular Biosciences, The Wenner-Gren Institute Stockholm University, SE-10691, Stockholm, Sweden.
Cancer Proteomics Mass Spectrometry, Department of Oncology-Pathology, Science for Life Laboratory, Karolinska Institute, Box 1031, SE-17121, Solna, Sweden.
PLoS One. 2014 Dec 31;9(12):e116294. doi: 10.1371/journal.pone.0116294. eCollection 2014.
A recently isolated phage, vB_EcoP_SU10 (SU10), with the unusual elongated C3 morphotype, can infect a wide range of Escherichia coli strains. We have sequenced the genome of this phage and characterized it further by mass spectrometry based proteomics, transmission electron microscopy (TEM), scanning electron microscopy (SEM), and ultra-thin section electron microscopy. The genome size is 77,327 base pairs and its genes, and genome architecture, show high similarity to the phiEco32 phage genes and genome. The TEM images reveal that SU10 have a quite long tail for being a Podoviridae phage, and that the tail also changes conformation upon infection. The ultra-thin section electron microscopy images of phages at the stage of replication within the host cell show that the phages form a honeycomb-like structure under packaging of genomes and assembly of mature capsids. This implies a tight link between the replication and cutting of the concatemeric genome, genome packaging, and capsid assembly. We have also performed a phylogenetic analysis of the structural genes common between Podoviridae phages of the C1 and C3 morphotypes. The result shows that the structural genes have coevolved, and that they form two distinct groups linked to their morphotypes. The structural genes of C1 and C3 phages appear to have diverged around 280 million years ago applying a molecular clock calibrated according to the presumed split between the Escherichia - Salmonella genera.
最近分离出的一种噬菌体,vB_EcoP_SU10(SU10),具有不寻常的细长C3形态类型,可感染多种大肠杆菌菌株。我们对该噬菌体的基因组进行了测序,并通过基于质谱的蛋白质组学、透射电子显微镜(TEM)、扫描电子显微镜(SEM)和超薄切片电子显微镜对其进行了进一步表征。基因组大小为77,327个碱基对,其基因和基因组结构与phiEco32噬菌体的基因和基因组高度相似。TEM图像显示,作为短尾噬菌体科的噬菌体,SU10具有相当长的尾部,并且尾部在感染时也会改变构象。宿主细胞内复制阶段噬菌体的超薄切片电子显微镜图像显示,噬菌体在基因组包装和成熟衣壳组装过程中形成蜂窝状结构。这意味着串联基因组的复制和切割、基因组包装和衣壳组装之间存在紧密联系。我们还对C1和C3形态类型的短尾噬菌体科噬菌体之间共有的结构基因进行了系统发育分析。结果表明,这些结构基因是共同进化的,并且它们形成了与各自形态类型相关的两个不同组。根据推测的大肠杆菌 - 沙门氏菌属之间的分裂校准分子钟,C1和C3噬菌体的结构基因似乎在约2.8亿年前就已经分化。