File:Ppat.1006562.g001.tif

English: Host and viral membrane fusion generates a tunnel though which viral DNA is ejected. A-G. PBCV-1-infected chlorella cells at 1.5–2 min PI were immobilized with HPF-FS and thick sections were analyzed by STEM tomography. A. A 5.2 nm tomographic slice from a 220 nm-thick STEM tomogram showing the close proximity between the viral and host internal membranes resulting from their convergence at the infection site. B. A 7.8 nm tomographic slice of a high magnification of the inset in panel A. C. A 5.2 nm tomographic slice from a different 220 nm STEM tomogram. D. High magnification of the inset of panel C. The generation of a continuous tunnel is evident. E, F. Two different 5.2 nm STEM tomography slices from the same tomogram showing the same PBCV-1-infected cells with almost completely empty capsids in which the membrane tunnel is still detected. G. A 5.2 nm tomographic slice from a 216 nm-thick STEM tomogram exhibiting an empty capsid attached near thylakoid membrane stacks (red arrowheads). In all panels the membrane tunnel and the protrusion of the host membrane are marked with blue and white arrowheads, respectively. Asterisk: viral DNA. H, I. Volume rendering representation of the STEM tomogram shown in panel A. The 3D surface representation highlights the barriers that viral DNA has to overcome to reach the host nucleus (including cell wall, plasma membrane, cytoplasmic vesicles, Golgi, and photosynthetic membranes that were not captured in this tomogram). A PBCV-1 virion is attached to the cell wall (brown). The host membranes as well as cytoplasmic vesicles are marked in blue. The capsid is depicted in yellow, the internal viral membrane and the membrane tunnel (arrowheads) are shown in blue. Viral DNA is shown in green. Scale bars: A, C, G: 100 nm; B, D, E, F: 50 nm.