ultrastructural characterization of the giant volcano-like virus factory of acanthamoeba polyphaga mimivirus超微结构特征的巨大volcano-like多食棘阿米巴巨型病毒工厂米米病毒.pdfVIP

Ultrastructural Characterization of the Giant Volcano-like Virus Factory of Acanthamoeba polyphaga Mimivirus Marie Suzan-Monti*, Bernard La Scola, Lina Barrassi, Leon Espinosa, Didier Raoult* ´ ´ ´ ´ ´ ´ Unite des Rickettsies, Centre National de la Recherche Scientifique (CNRS) UMR 6020, IFR 48, Faculte de Medecine, Universite de la Mediterranee, Marseille, France Acanthamoeba polyphaga Mimivirus is a giant double-stranded DNA virus defining a new genus, the Mimiviridae, among the Nucleo-Cytoplasmic Large DNA Viruses (NCLDV). We used utrastructural studies to shed light on the different steps of the Mimivirus replication cycle: entry via phagocytosis, release of viral DNA into the cell cytoplasm through fusion of viral and vacuolar membranes, and finally viral morphogenesis in an extraordinary giant cytoplasmic virus factory (VF). Fluorescent staining of the AT-rich Mimivirus DNA showed that it enters the host nucleus prior to the generation of a cytoplasmic independent replication centre that forms the core of the VF. Assembly and filling of viral capsids were observed within the replication centre, before release into the cell cytoplasm where progeny virions accumulated. 3D reconstruction from fluorescent and differential contrast interference images revealed the VF emerging from the cell surface as a volcano-like structure. Its size dramatically grew during the 24 h infectious lytic cycle. Our results showed that Mimivirus replication is an extremely efficient process that results from a rapid takeover of cellular machinery, and takes place in a unique and autonomous giant assembly centre, leading to the release of a large number of compl