National Center for Macromolecular Imaging, Verna and Marrs McLean Dept. of Biochemistry &Molecular Biology, Baylor College of Medicine, Houston, TX, 77030, USA.
National Institute for Physiological Sciences, 38 Nishigonaka, Myodaiji, Okazaki, Aichi, 444-8585, Japan.
Sci Rep. 2017 Mar 10;7:44176. doi: 10.1038/srep44176.
Marine cyanobacteria perform roughly a quarter of global carbon fixation, and cyanophages that infect them liberate some of this carbon during infection and cell lysis. Studies of the cyanobacterium Prochlorococcus MED4 and its associated cyanophage P-SSP7 have revealed complex gene expression dynamics once infection has begun, but the initial cyanophage-host interactions remain poorly understood. Here, we used single particle cryo-electron tomography (cryo-ET) to investigate cyanophage-host interactions in this model system, based on 170 cyanophage-to-host adsorption events. Subtomogram classification and averaging revealed three main conformations characterized by different angles between the phage tail and the cell surface. Namely, phage tails were (i) parallel to, (ii) ~45 degrees to, or (iii) perpendicular to the cell surface. Furthermore, different conformations of phage tail fibers correlated with the aforementioned orientations of the tails. We also observed density beyond the tail tip in vertically-oriented phages that had penetrated the cell wall, capturing the final stage of adsorption. Together, our data provide a quantitative characterization of the orientation of phages as they adsorb onto cells, and suggest that cyanophages that abut their cellular targets are only transiently in the "perpendicular" orientation required for successful infection.
海洋蓝藻进行大约四分之一的全球碳固定,而感染它们的蓝藻噬菌体在感染和细胞裂解过程中会释放部分碳。对聚球藻 MED4 及其相关的蓝藻噬菌体 P-SSP7 的研究揭示了感染开始后复杂的基因表达动态,但最初的蓝藻噬菌体-宿主相互作用仍知之甚少。在这里,我们使用单颗粒冷冻电子断层扫描(cryo-ET)技术,基于 170 次噬菌体-宿主吸附事件,研究了该模型系统中的蓝藻噬菌体-宿主相互作用。亚断层分类和平均显示了三种主要的构象,其特征是噬菌体尾部与细胞表面之间的角度不同。即,噬菌体尾部(i)与细胞表面平行,(ii)与细胞表面成约 45 度角,或(iii)与细胞表面垂直。此外,噬菌体尾纤维的不同构象与上述尾部方向相关。我们还观察到在垂直取向的噬菌体中,在穿透细胞壁后,在尾部尖端之外存在密度,捕获了吸附的最后阶段。总之,我们的数据提供了噬菌体在吸附到细胞时取向的定量描述,并表明与细胞靶标相邻的蓝藻噬菌体仅短暂处于成功感染所需的“垂直”取向。